Differentiation of rat iPS cells and ES cells into granulosa cell-like cells in vitro Juan Zhang, Hui Li, Zhao Wu, XiaoJun Tan, Fengying Liu, Xianghong Huang and Xiaoling Fang Acta Biochim Biophys Sin 2013, 45: 289–295; doi: 10.1093/abbs/gmt008 Department of Obstetrics and Gynecology, Second Xiangya Hospital, Central South University, Changsha 410083, China Premature ovarian failure (POF) is an ovarian defect characterized by the premature depletion of ovarian follicles before 40 years of age, representing one major cause of female infertility. Stem cells provide the possibility of a potential treatment for POF. In this study, rat embryonic stem cells (ESCs) and induced pluripotent stem cells (iPSCs) were co-cultured with granulosa cells (GCs) to differentiate to GC-like cells. The level of estradiol (E2) analyzed by radioimmunoassay showed that the E2 concentration of the culture supernatant of co-cultured rat iPSCs and ESCs increased in a time-dependent manner, compared with the GCs group that has an opposite trend. The expression of follicle-stimulating hormone receptor (FSHR) was confirmed by immunostaining. These results indicated that rat iPSCs and ESCs were effectively induced to GC-like cells through indirect cell-to-cell contact. Real-time polymerase chain reaction was performed to analyze the expression level of marker genes in POF, including BMP15, FMR1, FSHR, INHA, AMH, NOBOX, FOXO3, EIF2B, FIGLA, and GDF9. The BMP15, FSHR, INHA, AMH, NOBOX, and GDF9 genes were significantly up-regulated in iPSCs and ESCs co-cultured with GCs in comparison with cells that were not co-cultured. Thus, here we demonstrated an available method to differentiate rat iPSCs and ESCs into GC-like cells in vitro for the possible cell therapy of POF. Expression of FSHR in iPSCs and ESCs co-cultured with GCs 全文: http://abbs.oxfordjournals.org/content/45/4/289.full 相关论文: 1 Ovarian adult stem cells: hope or pitfall? 2 Efficient Differentiation of Steroidogenic and Germ-Like Cells from Epigenetically-Related iPSCs Derived from Ovarian Granulosa Cells 3 Generation of Germ Cells In Vitro in the Era of Induced Pluripotent Stem Cells 4 Efficient Induction of Pluripotent Stem Cells from Granulosa Cells by Oct4 and Sox2 5 Optimal vitrification protocol for mouse ovarian tissue cryopreservation: effect of cryoprotective agents and in vitro culture on vitrifiedwarmed ovarian tissue survival 6 Sex-determining region Y-box 2 and GA-binding proteins regulate the transcription of liver receptor homolog-1 in early embryonic cells 7 EDA-Containing Fibronectin Increases Proliferation of Embryonic Stem Cells
Dynamic changes in the copy number of pluripotency and cell proliferation genes in human ESCs and iPSCs during reprogramming and time in culture Free pdf http://acgt.cs.tau.ac.il/papers/PIIS1934.pdf Cell Stem Cell：ips细胞具更高基因畸变频率 2011-03-12 加州大学圣地亚哥分校医学院及斯克里普斯研究所的干细胞科学家领导的跨国研究团队，记录了在人类胚胎干细胞(hESC)和诱导功能干细胞(iPSC)系中特殊的基因畸变，题为《在细胞重组和培养过程中人类胚胎干细胞和诱导多功能干细胞的细胞增殖和全能性的拷贝数量上的动态变化》已在1月7日的Cell Stem Cell上发表。该公布的发现强调了需要对多能干细胞进行频繁的基因检测以保证其稳定性和临床安全性。 该研究的第一作者，加州大学圣地亚哥分校再生医学系的路易斯.劳伦特博士认为，我们发现人类多能干细胞(hESC和iPSC)比其他类型细胞有更高的基因畸变的频率。最令人吃惊的是，与其他非多能干细胞样本相比较，我们观察到hESCs的基因扩增和iPSC的缺失方面出现的频率更高。 人类多能干细胞在人体内具有发展成其他类型细胞的能力，可成为替换细胞治疗的潜在来源。斯克里普斯研究员再生医学中心主任，珍妮.罗伦教授谈到，由于基因畸变常常与癌症相关联，免受癌症相关的基因突变对于临床使用的细胞系来说至关重要。 研究团队确认了在多能干细胞系中可能发生突变的基因区域。对于hESC而言，可观察到的畸变大多是靠近多潜能相关基因区域的基因扩增；对于iPSC而言，扩增主要涉及细胞增殖基因及与肿瘤抑制基因相关的缺失。传统的显微技术，如染色体组型分析可能无法检测到这些变化。研究组使用一种高分辨率的分子技术，称为"单核苷酸多态性(SNP)"，能观察到人类基因组里一百多万个位点里的基因变化。 劳伦说，我们惊喜地发现在较短时间培养中的基因变化，例如在体细胞重编程为多能干细胞的过程以及在培养中细胞的分化过程。我们不知道这会有怎样的影响，如果有的话，这些基因畸变都会对基础研究或者临床应用的结果产生影响，对此应当深究。 劳伦总结到，该研究结果解释了有必要对多能干细胞培养进行经常性的基因监控，SNP分析仍不失人类胚胎干细胞和多能干细胞日常监控的一部分，但是这一结果提醒我们应当予以重视。 Cell Stem Cell doi：10.1016/j.stem.2010.12.003 Dynamic Changes in the Copy Number of Pluripotency and Cell Proliferation Genes in Human ESCs and iPSCs during Reprogramming and Time in Culture Louise C. Laurent, Igor Ulitsky, Ileana Slavin, Ha Tran, Andrew Schork, Robert Morey, Candace Lynch, Julie V. Harness, Sunray Lee, Maria J. Barrero, Sherman Ku, Marina Martynova, Ruslan Semechkin, Vasiliy Galat, Joel Gottesfeld, Juan Carlos Izpisua Belmonte, Chuck Murry, Hans S. Keirstead, Hyun-Sook Park, Uli Schmidt, Andrew L. Laslett, Franz-Josef Muller, Caroline M. Nievergelt, Ron Shamir, Jeanne F. Loring The tremendous self-renewal and differentiation capabilities of human pluripotent stem cells (hPSCs) make them potential sources of differentiated cells for cell therapy. Cell therapies are subject to rigorous safety trials, and high priority is placed on demonstrating that the cells are nontumorigenic (Fox, 2008). Because genetic aberrations have been strongly associated with cancers, it is important that preparations destined for clinical use are free from cancer-associated genomic alterations. Human embryonic stem cell (hESC) lines have been shown to become aneuploid in culture (Baker et al., 2007,Draper et al., 2004,Imreh et al., 2006,Maitra et al., 2005,Mitalipova et al., 2005), and the most frequent changes, trisomies of chromosomes 12 and 17, are also characteristic of malignant germ cell tumors (Atkin and Baker, 1982,Rodriguez et al., 1993,Skotheim et al., 2002). Aneuploidies can be detected by karyotyping, but less easily detectable subchromosomal genetic changes may also have adverse effects. Small abnormalities have been detected in hESCs by using comparative genomic hybridization (CGH) and single-nucleotide polymorphism (SNP) genotyping (Lefort et al., 2008,N?rv? et al., 2010,Spits et al., 2008). These studies lacked sufficient resolution and power to identify cell type-associated duplications and deletions. A recent study has reported the use of gene expression data to detect genomic aberrations in a large number of hESCs and hiPSCs (Mayshar et al., 2010). However, the methods used could reliably detect only relatively large (≥10 megabase) aberrations, and the lack of nonpluripotent samples for comparison precluded the authors from determining which regions of genomic aberration were specific to pluripotent stem cells. In this study, we performed high-resolution SNP genotyping on a large number of hESC lines, induced human pluripotent stem cell lines (hiPSCs), somatic stem cells, primary cells, and tissues. We found that hESC lines had a higher frequency of genomic aberrations compared to the other cell types. Furthermore, we identified regions in the genome that had a greater tendency to be aberrant in the hESCs when compared to the other cell types examined. Recurrent regions of duplication were seen on chromosome 12, encompassing the pluripotency-associated transcription factor NANOG and a nearby NANOG pseudogene, and on chromosome 20, upstream of the DNA methyltransferase DNMT3B. Although the frequency of genomic aberrations seen in the hiPSC lines was similar to those of cultured somatic cells and tissues, we observed one of the recurrent areas of duplication characteristic of hESCs in one of the hiPSC lines. Furthermore, comparison of 12 hiPSC lines generated from the same primary fibroblast cell line identified genomic aberrations that were present in the hiPSC lines and absent from the original fibroblast line. Analysis of early- and late-passage samples from these hiPSC lines allowed us to distinguish between events that arose during the process of reprogramming and those that accumulated during long-term passage. In general, deletions tended to occur with reprogramming and involve tumor-suppressor genes, whereas duplications accumulated with passaging and tended to encompass tumor-promoting genes. These results suggest that human pluripotent stem cell populations are prone to genomic aberrations that could compromise their stability and utility for clinical applications and that reprogramming and expansion in culture may lead to selection for particular genomic changes. 原文链接：http://www.medicalnewstoday.com/articles/213047.php
Nature 2011 Copy number variation and selection during reprogramming to pluripotency Free pdf http://proj1.sinica.edu.tw/~tigpcbmb/spring2011/Seminar/student/paper_YiHuiHsieh_2011.pdf http://www.39kf.com/focus/lc/clone/2011-03-19-711209.shtml Nature三篇文章：iPS细胞的遗传缺陷 来源：生物通 2011-3-19 2006年日本科学家山中申弥利用病毒载体将四 转录因子（Oct4，Sox2，Klf4和c-myc）的组合转入分化的体细胞中，使其重编程而得到了类似胚胎干细胞的 细胞类型——诱导 能干细胞（iPS cells） 种通过将完全分化的体细胞重编程，不经胚胎阶段而直接逆转至多能干细胞状态的iPS 细胞一度被视 最有希望运用到再生医学及新药开发的重要资源，为人类各种遗传性及功能性疾病的研究和治疗带来了新希望。然而近几个月来，陆续有一些研究团体报道发现iPS 细胞存在着重编程错误及基因组不稳定性的缺陷，这些缺陷将有可能导致iPS 细胞的临床治疗潜能受到限制。 近日国际著名生物学期刊《自然》（Nature）杂志同期发表了三篇论文分别针对iPS细胞的点突变、拷贝 变异及DNA甲基化特征进行了研究，新研究发现或将促使研究人员 审慎地利用iPS细胞开展干细胞研究及临床应用。 原文检索： Gore, A. et al. Somatic coding mutations in human induced pluripotent stem cells. Nature 471, 63–67 (2011) ArticleHussein, S. M. et al. Copy number variation and selection during reprogramming to pluripotency. Nature 471, 58–62 (2011) ArticleLister, R. et al. Hotspots of aberrant epigenomic reprogramming in human induced pluripotent stem cells. Nature 471, 68–73 (2011) 在第一篇文章中，Gore等针对来自7个实验室利用5种不同方法诱导人类成纤维细胞生成的22种iPS细胞系进行了外显子组测序。研究人员在这些iPS细胞中确认了124个点突变，由此他们推测出每个iPS细胞基因组中约存在6个编码区突变。值得注意的是，他们鉴别了多个有可能改变蛋白质功能的错义突变，同时发现一些点突变富集于与癌症相关的基因中。研究人员利用超深度测序（ultradeep sequencing）证实约一半的突变以低水平存在于起源的成纤维细胞群中，而另一半则发生在重编程过程中或之后。作者们认为这些突变负荷有可能是在重编程和培养过程中受到了选择作用而并非重编程固有诱变作用的影响。 在第二篇论文中Hussein及同事研究了iPS细胞的基因组完整性。利用一种高分辨单核苷酸多态性（SNP）分析技术平台，他们比较了 量人类胚胎干细胞(ESC)系、人类iPS细胞系以及起源的成纤维细胞系之间存在的拷贝数变异（CNV）的差异。研究人员证实相对于胚胎干细胞，iPS细胞有着更多的拷贝数变异，然而随着细胞传代这些变异数量逐渐减少。作者们证实在重编程过程中高比例地形成CNVs导致了早期几代iPSC细胞群中的遗传镶嵌现象。在细胞增殖过程中，快速选择淘汰了具有大量CNVs的细胞，从而使得iPSCs在晚期几代中逐渐与胚胎干细胞相似。然而，作者谨慎地表示某些CNVs有可能为细胞提供了选择利益。 在第三篇论文中，Lister等利用鸟枪法测序技术以单碱基分辨率在iPSCs, ESCs,体细胞以及由多能细胞分化的细胞中进行了全基因组DNA甲基化测定。研究人员发现尽管iPSCs和ESCs的甲基化组存在大量相似之处，然而研究的5个iPS细胞系与ESCs还是有着显著的差异，某些差异甚至在分化后持续存在。而iPSCs之间的差异则体现在细胞中的甲基化标记上,这些甲基化标记代表了起源的体细胞类型的“记忆”效应以及其他一些iPSC特异性改变。值得注意的是这些细胞系之间显示出了差异的甲基化区域（表明某些区域尤其易于发生异常的重编程）。在着丝粒和端粒附近的巨碱基区域显示了非CG序列甲基化改变，与转录及组蛋白甲基化中的改变相关。 除了强调研究人员需要更仔细地鉴定iPS细胞系的特征，这些研究还针对细胞分裂和培养的相对影响，以及重编程中固有的分子事件提出了一些有趣的问题。 （生物通：何嫱） 延伸阅读: • 最新Nature文章说说iPS的缺陷 来自哈佛干细胞研究院，霍德华休斯医学院等处的研究人员在实验中发现诱导多能干细胞（iPS）在分化发育方面的能力不及胚胎干细胞，研究人员还找出了造成这一情况的原因之一。这一研究成果公布在4月25日的Nature杂志上。文章的通讯作者是来自哈佛大学的KonradHochedlinger博士，这位著名的iPS研究人员在诱导多能干细胞研究领域获得了许多重要的成果，比如他曾培育出没有永久性基因损害的iPS，而这种以往的基因损害与人们的设计有关。这一发现代表了人们在未来临床上应用iPS细胞迈出了重大的一步。iPS技术利用不会自身被整合到宿主基因组内的病毒，诱导形成多能干细胞，绕过了使得以往iPS细胞的成功遭... 详细 • p53，iPS技术登Nature研究亮点 2007年自然出版集团宣布《Nature》出版集团的新出版物、名为NatureChina的网站（www.naturechina.com.cn）正式启动。这一网站致力于聚焦中国大陆地区和香港的优秀科学成果，每周都会针对最新发表的论文，在此网站撰写摘要和评述。本期的推荐研究文章包括：WinterTemperatureandUVAreTightlyLinkedtoGeneticChangesinthep53TumorSuppressorPathwayinEasternAsia据昆明动物所报道，研究结果发现在东亚人群中，p53基因第72位的序列变异(脯氨酸变为精氨酸)同纬度密切相关，即纬度越高，其所在... 详细 Blog， 公司不能打开： 2. 顶奸杂志《自然》开始报告IPSC的黑暗面_刘实_新浪博客 blog.sina.com.cn/s/blog_502041670100pcx7.html - 网页快照 2011年3月3日 – Copy number variation and selection during reprogramming to pluripotency. • Samer M. Hussein,. • Nizar N. Batada,. • Sanna Vuoristo, ...
2012-10-31 20:57:09 来源：光明网卫生频道 刘实 查看评论 进入光明网BBS 手机看新闻 !--enpproperty 55407532012-10-31 20:57:09.0刘实《科学》网站登刘实就iPS研究获诺贝尔奖致山中伸弥的公开信(1)_专家·专栏 _光明网细胞,多能干细胞,诺贝尔奖29201专家·专栏/nodesearchname/enpproperty-- 刘实 10月30日《科学》网站发表新闻 “Japanese Paper That Fell For False Stem Cell Claim Takes Corrective Action” （日本报纸因登虚假干细胞成果而采取纠错行动）。 在该新闻下。刘实贴出了他今日发给iPS细胞之父、今年的诺贝尔医学奖得主之一的日本人山中伸弥的公开信。 【背景】 2006年日本科学家山中伸弥在《细胞》杂志发表了第一篇关于iPS细胞的论文，同期刊登的一篇评论文章称山中伸弥实现了将末端分化的成年体细胞转化为胚胎干细胞样的多能干细胞，也就是大众媒体传说的“返老还童”。 对于这一惊人的声称，当时科学界都持谨慎态度，因为大家还没从韩国黄禹锡干细胞造假的阴影走出。 2007年《自然》杂志发表山中伸弥的第二篇iPS细胞论文，同期发表的另一iPS细胞论文则是美国科学大腕“夜里喜”（Jaenisch的音译）团队做出的。而当年创刊的《细胞-干细胞》杂志也发表了“夜里喜”以前的学生所做的另一个iPS细胞研究论文。这三篇论文的同时发表立刻造成了轰动效应，令科学界的主流不得不折服iPS重编程确有“返老还童”的魔力。 然而，对生命本质及发育过程有独到见解的刘实（ 详情 和 详情2 ）对这些iPS细胞论文所声称的将末端分化成年体细胞转化为与胚胎干细胞不可区别的多能干细胞表示怀疑，并明确指出所谓的iPS细胞就是错编程的干细胞或者说是人造癌细胞。这样的细胞不是不可与胚胎干细胞相区别，而是肯定有区别。这样的细胞不是安全和道德的胚胎干细胞替代品，用他们做细胞疗法是有致癌危险的（ 详情 ）。 山中伸弥在2007年答复刘实投给《自然》的批评文章时斩钉截铁地说他从未声称过将末端分化的成年体细胞转化为胚胎干细胞样的多能干细胞，并同意刘实提出的iPS细胞可能来自于本身就存在的干细胞或前细胞的观点。山中伸弥于2008年在《科学》发表更正时更是点名感谢刘实对其发表在《科学》的“无癌症风险”iPS细胞论文的批驳。 刘实对iPS细胞研究中表现出的一系列伪科现象的批驳发表在众多的论文中，其中有世界第一篇经过同行评审而于2008年发表在干细胞研究老牌主流杂志《干细胞与发育》上的综述。该综述发表后成为该杂志下载量最多的论文，两年之内被下载6500余次。 从2008年开始，刘实每年都给诺贝尔奖评委会发信，揭示iPS细胞研究的虚伪面并要求评委排除影响因子的影响而不错奖一个子虚乌有的发现。因为最新的实验报告实际上已完全证明刘实关于iPS细胞就是将干细胞错编程为人造癌细胞的基本判断，刘实今年更是两次给诺贝尔奖评委会发信。 然而，一个诺贝尔奖的悲剧还是发生了：山中伸弥竟因“发现成熟细胞可被重编为多能”而得了2012年的诺贝尔医学奖。对此，刘实已在《科学》网站发表多篇评论，认为诺贝尔奖被错误地发给了一个想像中的发现。 现在，刘实更是直接致（电子）信给山中伸弥，要求他明确答复他是否真的实现了将末端分化的成年体细胞转化为胚胎干细胞样的多能干细胞，同时请山中伸弥去斯德哥尔摩领奖前思考并回答一些关键问题。 刘实给山中伸弥的信现还以公开信的方式发布在《科学》的网站上。全文如下： 刘实 Open Letter to Yamanaka on His Nobel Prize-Winning iPS Work 就诺贝尔奖给iPS研究致山中伸弥的公开信 October 30, 2012 2012年10月30日 Dear Dr. Yamanaka, 亲爱的山中博士， As a solid scientist who was once publicly thanked by you in Science for correctly criticizing your invalid claim of cancer-free iPS cells, I am writing to you again to repeat my challenge to your now accepted claim of inducing terminally differentiated cells into pluripotent stem cells that are indistinguishable from embryonic stem cells (ESCs). 作为一个曾因正确批评你无效的“不致癌iPS细胞”而被你在《科学》点名公开感谢的扎实科学家，我再次给你写信重复我对你现已接受的诱导末端分化细胞为与胚胎干细胞不可区别的多能干细胞的声称的挑战。 In response to my Communications Arising questioning the origin of iPS cells reported on your 2007 Nature paper you stated that “We agree that the origin of iPS cells may be tissue stem or progenitor cells co-existing in fibroblast cultures”. You even declared that“We have never claimed that we generated iPS cells from terminally differentiated cells.” Furthermore, with reference to my proposed single-cell tracking experiments, you stated that “I agree that the experiments you proposed are important.” 在回复我对你2007年《自然》论文的iPS细胞来源的质疑时，你说“我们同意iPS细胞可能起源于纤维母细胞培养中共存的组织干细胞或前细胞”。你甚至于宣布“我们从未声称过从末端分化细胞产生iPS细胞”。而且，针对我提出的单个细胞跟踪实验，你说“我同意你提出的实验是重要的”。 Now, a Nobel Prize was awarded to you “for the discovery that mature cells can be reprogrammed to become pluripotent” which in fact refers to a claim made for you that you have advanced “a significant step toward delineating the minimal set of factors required to confer the developmental potential of an embryonic stem(ES) cell onto a terminally differentiated somatic cell” or, in popular term, you have found a “simple switch turns cells embryonic”. 现在，一个诺贝尔奖因为你“发现成熟细胞可被重编为多能”而奖给了你。这实际上是相信了一个为你制造的声称：你在阐明赋予末端分化体细胞以胚胎干细胞样发育潜能的最低因素上迈了一个显著的步伐。用通俗的话说，你发现了将细胞变为胚胎样的简单开关。 Apparently, you are very happy to accept these assessments and most likely will go to Stockholm to accept the Nobel Prize. But, before you do that, you need to think carefully the following points: 很明显，你是非常乐意接受这些评价而且很可能会去斯德哥尔摩领诺贝尔奖。但在你做那之前，你需要仔细考虑下面几点： First, are you sure now you have achieved reprogramming terminally differentiated non-stem adult/mature cells into ESC-like pluripotent stem cells? If so, what are the evidences you have reported for that claim? 第一，你现在可明确你实现了重编末端分化非干细胞的成年/成熟细胞为胚胎干细胞样的多能干细胞吗？ Second, have you done the single-cell tracking experiments that I proposed to you back in 2007 and were considered as important by you then? If yes, what are the results? If not, why not? 第二，你做了我2007年就给你提出的而你也认为重要的单个细胞跟踪实验吗？如果做了，结果如何？如果没做，是因什么？ Third, what is your opinion about Jaenisch group’s new findings that only a very small fraction (0.01%) of donor cells can be turned into (fully) reprogrammed iPSCs and, more significantly, iPSCs can be sorted out according to some stemness markers without using any generic “Yamanaka” factors? 第三，你怎么看待“夜里喜”团队的新发现：只有很小一部分（0.01%）供体细胞可被变为完全重编程的iPS细胞。更为重要的是，不用标准的“山中”因子也可根据干性标志筛选出iPS细胞。 Fourth, do you still think that iPS cells are not intrinsically cancerous cells and thus can be safely used for achieving cancer-free cell therapy? 第四，你仍然认为iPS细胞不是本能的癌化细胞而且可被安全地用于无癌发生的细胞疗法吗？ Fifth, do you still think that the observed rejection of syngenic iPS cells by animals with normal immunity is a questionable finding and syngenic iPS cells should have a natural advantage against immune rejection? 第五，你仍然认为被观察到的免疫正常的动物排斥自体来源的iPS细胞是可疑的发现而且自体来源的iPS细胞就应当具有天然的避免免疫排斥的优点吗？ I think your answers to above questions will help you to reach a safe conclusion on whether or not you should accept the Nobel Prize. 我想你对上述问题的回答将有助于你做出一个是否接受诺贝尔奖的安全结论。 Honestly, if I were you, I will ask the Nobel Prize Assembly to delay the award until I have obtained at least some solid evidence that I can indeed reprogram any mature/adult differentiated non-stem cells back into ESC-like cells. 老实地说，如果我是你，我会要求诺贝尔大会推迟发奖，直到我至少获得了一些扎实的证据表明我确实能够重编任何成年/成熟的非干细胞为胚胎干细胞样的细胞。 Sincerely, 诚挚的， Shi V. Liu 刘实 附： Japanese Paper That Fell For False Stem Cell Claim Takes Corrective Action byDennis Normileon30 October2012,10:55 AM| The Japanese newspaper that published what turned out to be false newsof the first clinical application of cells derived from induced pluripotent stem cells has detailed the lessons it learned and the steps it is taking to prevent a recurrence. The Yomiuri Shimbun,Japan's largest circulation daily, already apologizedto readers for what it admitted were a collection of false articles. Now, in a collection of articles in its 26 October Japanese morning edition and the 27 October English Daily Yomiuri(available online here, here, here, and here), the media organization explained that reporters and editors failed to check basic facts. The newspaper detailed punitive pay cuts for those involved and the replacement of the science news editor. To head off such problems in the future, the paper pledged to thoroughly check facts, strengthen the verification system in the science news department, and foster the development of reporters who are “able to put matters in perspective." TheYomiurialso explained that its staff members had failed to check previous statements made by Hisashi Moriguchi, the man at the center of the scandal, that were reported in a number of previously published articles. These included claims of an affiliation withHarvardMedicalSchooland of research accomplishments. This reporting gave Moriguchi, "a false ladder to academic success," the English language article states. 点击进入： 刘实 专家专栏
2012 Nobel Prize in Physiology or Medicine The 2012 Nobel Prize in Physiology or Medicine was awarded jointly to John B. Gurdon and Shinya Yamanak a " for the discovery that mature cells can be reprogrammed to become pluripotent ". 2012年诺贝尔生理或医学奖被授予了英国的 John B. Gurdon 和日本京都大学科学家 Shinya Yamanak a，因他们发现成熟的体细胞可以被重编程具有多能性。 他们的贡献呢？ “1962 年，29岁的牛津大学科学家的John Gurdon将发育到了蝌蚪阶段的非洲爪蟾肠细胞的 核移入到已移除细胞核的非洲爪蟾受精卵中，获得了很多具有相同基因背景的 非洲爪蟾——它们都来自于提供肠细胞的蝌蚪 (1.Gurdon, 1962)。这提示了动物 细胞的体细胞依然具有发育为一个完整个体的全能性。 2006 年，日本科学家山中伸弥 （Shinya Yamanaka）通过逆转录病毒导入 四个转录因子（Oct4, Sox2, Klf4, c-Myc, SKOM，四因子，4F；我们也将前三个 转录因子称为三因子，3F）直接将鼠胚胎纤维原细胞（Mouse embryonic fibroblast, MEF）和鼠尾尖细胞（mTTF）转变到了一个诱导的多能干细胞状态 (2.Takahashi and Yamanaka, 2006)。这两位科学家因为以上工作共同获得了2009 年的拉斯克 医学研究奖，山中伸弥还在《自然医学》杂志上发表的获奖感言——《iPS细胞 的漫漫接力长路》 (3. Yamanaka, 2009)中特意提到和Gurdon的缘分，山中出生的 1962 年正是Gurdon做出核移植爪蟾的那年。” 现在他们幸福地一起过着诺贝尔奖得主的生活。 Papers： 1 Gurdon JB. Adult frogs derived from the nuclei of single somatic cells. Developmental Biology 1962; 4 (2): 256-73. 2 Takahashi K, Yamanaka S. Induction of pluripotent stem cells from mouse embryonic and adult fibroblast cultures by defined factors. Cell 2006; 126 (4): 663-76. 3 Yamanaka S. Ekiden to iPS Cells. Nat Med 2009; 15 (10): 1145-8. eng. 酷酷的发型 NOBEL网站的黑板报已经很多人赞他的发型了。图片来自诺贝尔奖网站。
1. yamanaka缺乏科学实验基本功，所用供试样品细胞未经过分离纯化，是混杂的细胞群，还包括什么muse之类的... 2. 欺世盗名，把人造畸形细胞强说成是类胚胎干细胞：iPS仅仅激活了部分全能性基因，并未对起始细胞的表观遗传特征进行抹除，有意忽略iPSC的表观遗传缺陷... 3. 所谓“返老还童”根本没实现，全能性验证试验具有欺骗性，硬是把胚性微环境中表观遗传的重构说成是iPS的全能性... 4. 哗众取宠，任何有常识的人都知道转分化是条捷径，而yamanaka偏偏多此一举搞所谓“拨回发育时钟”... 5. Yamanaka从未作出过真正的重编程多能干细胞（iPSC），只是硬着头皮把“转基因技术”说成是“细胞重编程”... 6. 和黄禹锡如出一辙，骗取经费； 7. Yamanaka很具有日本人的传统性格，宁死不认错 ！ 【小资料】诱导多能干细胞（induced pluripotent stem cells, iPS cells）最初是日本人 山中申弥 （Shinya Yamanaka）于2006年利用 病毒载体 将四个 转录因子 （Oct4, Sox2, Klf4 和c-Myc）的组合转入分化的体细胞中，使其 重编程 而得到的类似 胚胎干细胞 的一种细胞类型。2007年、2008年和2010年美国《科学》杂志都以iPS细胞倒转“生命时钟”为名将其评选为年度十大科学突破。 【名词解释】 细胞重编程（Reprogramming ） ：对已分化细胞的表观遗传特征抹除和重构（ refers to erasure and remodeling of epigenetic marks, such as DNA methylation , during mammalian development）。 转基因技术（Transgene）：通过自然或其它基因工程手段将基因及遗传物质从一种生物体导入到生另一个生物基因组中（a gene or genetic material that has been transferred naturallyor by any of a number of genetic engineering techniques from one organism to another）； 【寓言故事一】 “重编程”or“转基因”？ 日本男孩Yamanaka偷了邻家的四粒花种（基因：Oct4, Sox2,c-myc 和 Klf4），拌上肥料撒到自家园子里（转基因），花园里长出了原先没有的新花品种（所谓iPSC），邻家找上门来质问，Yamanaka辩称：我只是给花园施了些肥（重编程）； 【寓言故事二】中国男孩指着iPSC问yamanaka说：这家伙怎么不穿衣服？ 【魔鬼词典】重编程多能干细胞（iPS cell）是被转基因、转RNA、转蛋白、转小分子、吃Vc、乏氧等折腾得想让干什么就得干什么的“老顽童”细胞； 【疑问】教父级人物弄出如此低级的错误是主观故意还是客观无能？iPS的概念有无学术底线？
ScIence 和 Nature 在国人心目中是具有同等的至高无上地位的圣刊，历年年底他们都要评出当年科学上的突破性进展，也就必然引起国人的瞩目，甚至会影响国家有关科学研究基金的导向。今年 Science 打破历年的惯例，不再闭门造车，而是要广开言路，听取科学家的意见，也算给我们这个被认为缺少民主的国家做做示范。 2008 年 Science 曾把诱导性多能干细胞（ IPS ）的研究推为当年的突破性进展，老朽曾因根据本人在植物中的研究，在本博客发过几篇对此类研究的评论性博文。所以对此问题一直很关心。另外虽对刘实这个科学狂人批评过，但对他的洞察力还是很欣赏很佩服的。而且我在植物中的科研结论与他的一些科研成果又有许多正好互相印证的不谋而合，所以也一直很关注他对 IPS 的批评。恰好，看到 Cell Stem Cell 发的证明诱导性多能干细胞在培养条件下发生染色体异常变化的论文及 Cell 发的相关新闻评论。这就使我想到这进一步证明了刘的有关文章。 12 月 9 日 看到 Science 就评选 2010 年科学突破性进展征求意见后，就写了篇《 Reprogramming out of the iPS detour 》，希望不要再将此类研究作为突破性进展。谁知此后不断有人写意见推荐刘实的有关发现。 12 月 13 日 一个署名 Adam 的以给 Science 编辑信的形式发表意见，说刘实的研究只是科学的侧流，不应该让侧流干扰主流科学，应把提名刘实的评论统统删掉。我看到后很生气，当即也给编辑写了信（没认真考虑和修改，英文有些蹩脚），亮明我自己的身份，问 Adam 是谁？指出如果编辑部只想听自己喜欢的意见，不想听取不同的意见，就将我们的意见删除。此后还是不断有提名刘实的意见发表。 12 月 14 日 编辑部以 Science Staff 的名义声明，欢迎每一个人参加讨论，希望保留每个人的意见。但此后就再也看不到新的意见发表。于是我就又写了篇题目为 The Breakthrough of the Year in Science: Does iPS stand for incorrect programming syndrome? 的评论意见想在今天放上去，但当发布时却显示出，每个评论不能超过 300 字，于是我就改成了 3 篇重发。这次虽发出了，但文后却出现了一行字 Message awaits moderator approval ，也就是说要经过管理人员审查后才能发布，这在以前是没有的。但等今天下午我再进去看时，不仅发评论的窗口不见了，连所有意见也不见了。看来是停止听取意见了，为什么不预告呢，这是人家的自由，人家的权利。 看来所有的民主和自由都不是不受限制的，连讨论科学上的突破性进展都如此，还用说其他的吗？美国的民主自由，英国的民主自由都有自己的标准，为什么别的国家非要按他们的标准呢？ 附件是 Science 征求 2010 年科学突破性进展意见的专栏和我所记录下的所有评论的复印件，最后是我的三个没正式发布的评论。因我记录的最早的评论是 2010 年 12 月 6 日 ，故将题目定为 10 天民主。 Reader Opinions Name the 2010 Breakthrough by Science Staff | 60 comments (http://talk.sciencemag.org/nodes/btoy2010.html?s=ghp) Email Print | More Every December, our editors and News staff face the challenge of reviewing what science has accomplished around the world in the past 12 months, so as to select our breakthroughs of the year. The task is an invigorating one, providing a powerful reminder of both the enormous scope and the continual advance of science. Ardipithecus ramidus, reprogramming cells, and human genetic variation topped our lists in recent years. Now it's your turn: What would you dub the major scientific breakthrough of 2010? We're eager to hear your thoughts and will be sharing our picks with you soon! Comments ( Commenting Terms and Conditions ) These postings do not necessarily represent the views/opinions of Science . 2010/12/15 15：00成为空白 Keming Cui I say iPS is incorrect programming syndrome because it is indeed a syndrome of irresponsible research. As a botanist studying developmental biology for a life time and thus know some botanical cloning and reprogramming (if I can use these modern terms to characterizing our traditional studies) I found it is truly amazing that someone would rather call an neoplastic transformation as a regenerative reprogramming and regard cancerous cells as therapeutic cells. Fortunately, science is about searching for truth and truth will prevail eventually. Many studies revealing the true nature of iPS cells and iPS reprogramming have been published this year. Collectively these studies supported Dr. Lius discoveries on iPS cells and iPS reprogramming, even though they generally do not cite Dr. Lius PUBLICATIONS. Why? How could they do so? How could the journals allow them to do so? Finding out answers to the above questions may lead to a more profound breakthrough in science, a breakthrough lead to the establishment of responsible research. However, this breakthrough may not come this year but no one can stop its coming in the future. Period! Message awaits moderator approval 2010/12/15 , 9:02:45 Keming Cui We owe Shi V. Liu a big deal in breaking through iPS hype I have suggested that diagnosing iPS as incorrect programming syndrome is a breakthrough in science. Now I wish to say that it is not easy to reach this diagnosis which may still be refused by some iPS patients. However, history will show that diagnosis is absolutely correct and we owe Shi V. Liu a big deal in breaking through the iPS hype. Comparing what Dr. Liu has stated in his pioneering PUBLICATIONS and what others have discovered later, I found that Dr. Lius DISCOVERIES are not only very correct but also more insightful than all other studies reported in top journals! If Science Staff do not believe my assessment please spent a few days to read Dr. Lius PUBLICATIONS. I knew his PUBLICATIONS can be obtained free of charge and many people have already known his views on iPS in specific and cell life in general. Thus, if Science Staff still neglect Dr. Lius discoveries and even go ahead further promote flawed iPS research, a future breakthrough in science may come against Science when responsible research is demanded at all the levels, which I will continue in my part three of this long essay. Message awaits moderator approval 2010/12/15 ,9:02:07 Keming Cui The Breakthrough of the Year in Science: Does iPS stand for incorrect programming syndrome? Dear Science Staff, I agree with you 100% on your short but very clear request that contributors should keep their comments relevant to the topic of this year's breakthrough. What is this years breakthrough? What can be this years breakthrough as compared with or connected with previous breakthroughs? Let us recall what breakthroughs were made in Science recently. On the left side of this comment collecting window I saw 2008 Breakthrough of the Year: Reprogramming Cells. Apparently it refers mainly to the so-called discovery of iPS cells or iPS reprogramming. What are iPS cells and iPS reprogramming? Shi V. Liu concluded (as evidenced in many PUBLICATIONS collected at http://im1.biz/iPS.htm ) that iPS cells are NOT induced pluripotent stem cells but incorrectly programmed stem cells or man-made cancer stem cells. Thus, iPS reprogramming is essentially an incorrect programming syndrome. If that diagnosis is correct (which will be further argued in my next comment because of the space limitation here), then the CONFIRMATION of Lius discovery by others in this year is really a very relevant breakthrough in science for Science. Message awaits moderator approval 2010/12/15 , 9:01:31 標記 Science Staff We thank everyone for their contributions to this discussion, but urge people to please keep their comments relevant to the topic of this year's breakthrough. Thank you. 2010/12/14 , 8:04:38 Shapiro Another Set of Breakthrough Discoveries Arranged by Time For long time DNA/chromosome segregation has been regarded as a random process. However in early 1990s Dr. Liu has proposed that DNA/chromosome segregation should follow a regular pattern in that the older DNA template strand/chromosome is retained by the true mother cell while the younger DNA template strand/chromosome is distributed into the true daughter cells. He defines the true mother cell as the cell which has reproduced a daughter cell and often remains live to reproduce more daughter cells, base on his continuous tracking of individual bacteria and his pioneering discovery on bacterial cell life. However, Dr. Lius discovery was rejected for publication in many western journals he tried. Nevertheless his succeeded in publishing his discovery on bacterial life and also the above DNA/chromosome segregation hypothesis in 1999 Science in China. He further interpreted his findings in a series articles published in Logical Biology and, more importantly, in a patent application filed at 2000 and granted in 2004 by US Patent and Trademark Office. In 2005 Dr. Liu submitted to Nature a Hypothesis type article titled as Linking DNA aging with cell aging and combining genetics with epigenetics. Using in silico labeling, Dr. Liu depicted in very details the pattern of DNA/chromosome segregation during cell reproduction and also insightfully delineated the respective contribution of genetic and epigenetic factors to the inheritance and adaptation. Liu published this paper in Logical Biology (5:51-55, 2005) after rejection by Nature. Later Dr. Liu also published a paper entitled A theoretical framework for understanding biotic aging from molecule to organism in multicellular life (Logical Biology 5:109-116, 2005) which will prove to be one of his most significant publications and a historically landmark paper in biology. Tracking down later publications in mainstream journals I found that Dr. Lius discoveries very solid. These can be proved by comparing some high-profile publications appeared in some top journals including Nature, for examples: Armakolas, A., and A. J. Klar. 2006. Cell type regulates selective segregation of mouse chromosome 7 DNA strands in mitosis. Science 311:1146-9. Conboy, M. J., A. O. Karasov, and T. A. Rando. 2007. High incidence of non-random template strand segregation and asymmetric fate determination in dividing stem cells and their progeny. PLoS Biol 5:1120-1126 Falconer, E. et al. 2010. Identification of sister chromatids by DNA template strand sequences. Nature 463: 93-97. However, my view may be biased. Thus, I encourage people to read these publications and compare them with Lius earlier publications and then voice some opinions here. 2010/12/14 , 5:42:14 dxyue After reading the references posted in the previous comments, I believe that Dr. Shi Liu should be given priority and be recognized for his pioneering work, which was published as early as 1999 then confirmed by Ackermann et al. 2010/12/14 , 3:39:09 HuaKe I am a scientific friend of Dr. Liu since 1983 when we were graduate classmates and know many detailed sides of his discoveries and, more importantly, his frustrations in communicating his discoveries to the mainstream. After decade of rejections by western journals of his fundamental discovery in life science - the breaking of the cell division dogma - he followed my advice by seeking review from a different peer group - the Chinese scientists in the mainland of China. His paper, based on ones rejected by Science, Nature, and PNAS, was reviewed in China for over one and half year and then accepted. After the acceptance of his English version, the editor invited him to translate the accepted English version into Chinese. Thus, his landmark paper on bacterial life was published in both English and Chinese in 1999 in Science in China - Life Science. That discovery, in my view, deserves a Nobel Prize. To further communicate his discovery to wide readership and to promote freedom and constructive debate in scientific communication Dr. Liu launched the world-first (I believe) open-access scientific journal that is also open for unrestricted post-publication peer and public review and comment. He named the journal as Logical Biology because he realized that many biologists have been occupied with tiny details of data but lost big picture of logic and thus often see trees but lose sight of the forest. He published a Correspondence in Nature (I forget the citation for it) to make this revolution in scientific publishing known by outside world. Later I actually saw some other scientists published their often ground-breaking discoveries in Logical Biology. I think Science has done a great thing by asking opinions on nomination of the Breakthrough of the Year. I just wish that the actual selection process will indeed consider the opinions expressed by the various readers. Let us hope a real breakthrough will indeed happen in Science! 2010/12/14 , 1:13:01 Ray ZHANG I belive that tumors grow their own blood vessels is false !....the vessels may be derived from the residual normal stem cell in the tumors? 2010/12/13 , 23:16:28 UFO Revolution in scientific publishing yields fruitful results Recently I read some articles on open review in some traditional journals. However, as far as I know, a double open (open-access and open-review) scientific publishing model has been implemented by Logical Biology a decade ago ( http://logibio.com ). Actually, there was a Correspondence by Shi V. Liu in Nature (403: 592, 2000) which specifically call for dramatic change in scientific publishing. Ten years have passed. I found Logical Biology and other Truthfinding Cyberpress (TFCP) journals ( http://im1.biz ) actually published many ground-breaking discoveries that were only being confirmed in some incomplete fashions by publications in some traditional top journals. TFCP journals also published a lot of insightful articles criticizing the scientific mistakes and ethical problems in publications appeared in other journals, often after the rejections of these criticisms by the other journals. If one pays respect to true discoveries and genuine knowledge one may find that the best science may be found in some sidestream of scientific publishing. Seeing is believing. Go to some revolutionized publication platforms for scientific communications and to see the true front-line of scientific research. 2010/12/13 , 12:21:52 新潮 Revolution in scientific publishing yields fruitful results Recently I read some articles on open review in some traditional journals. However, as far as I know, a double open (open-access and open-review) scientific publishing model has been implemented by Logical Biology a decade ago ( http://logibio.com ). Actually, there was a Correspondence by Shi V. Liu in Nature (403: 592, 2000) which specifically call for dramatic change in scientific publishing. Ten years have passed. I found Logical Biology and other Truthfinding Cyberpress (TFCP) journals ( http://im1.biz ) actually published many ground-breaking discoveries that were only being confirmed in some incomplete fashions by publications in some traditional top journals. TFCP journals also published a lot of insightful articles criticizing the scientific mistakes and ethical problems in publications appeared in other journals, often after the rejections of these criticisms by the other journals. If one pays respect to true discoveries and genuine knowledge one may find that the best science may be found in some sidestream of scientific publishing. Seeing is believing. Go to some revolutionized publication platforms for scientific communications and to see the true front-line of scientific research. 2010/12/13 12:17:04 訪客 Revolution in scientific publishing yields fruitful results Recently I read some articles on open review in some traditional journals. However, as far as I know, a double open (open-access and open-review) scientific publishing model has been implemented by Logical Biology a decade ago ( http://logibio.com ). Actually, there was a Correspondence by Shi V. Liu in Nature (403: 592, 2000) which specifically call for dramatic change in scientific publishing. Ten years have passed. I found Logical Biology and other Truthfinding Cyberpress (TFCP) journals ( http://im1.biz ) actually published many ground-breaking discoveries that were only being confirmed in some incomplete fashions by publications in some traditional top journals. TFCP journals also published a lot of insightful articles criticizing the scientific mistakes and ethical problems in publications appeared in other journals, often after the rejections of these criticisms by the other journals. If one pays respect to true discoveries and genuine knowledge one may find that the best science may be found in some sidestream of scientific publishing. Seeing is believing. Go to some revolutionized publication platforms for scientific communications and to see the true front-line of scientific research. 2010/12/13 , 12:16:14 Dr. Honest Some people have expressed their support for nominating Dr. Shi Lius fundamental discovery in cell biology as the Breakthrough of the Year. I think, at the science level, all of the mentioned biology discoveries made by Liu are outstanding and have far-reaching implications. However, as a clinician, I felt most excited by Dr. Lius discovery on cancer, especially his unique view on the multigenesis of multisite cancer. This is because it has profound implications with the treatment options for the patient and may change the outcome of the therapy. The conventional view of regarding most if not all the multisite cancer as a result of metastasis has mandated aggressive whole body chemotherapy or radiation therapy once a primary (may not be true primary) cancer is found. However, our direct observations and some studies have shown that the outcome of patients accepting such treatment may not be better than those who did not go through this harsh treatment. If the (primary) cancer first detected is not the source of future spreading but an early and easy detection of some multigenerated cancer which may be rooted to some mutations in stem/progenitors cells already scattered in the different body parts over the development, then we really need to think what would be the best way to treat cancer patients. One thing that we have learned now is that cancer stem cells often resist conventional chemotherapy and can even come back more aggressively once the normal cells/immune system is jeopardized by the non-discriminating killing of all reproducing cells by the concessional chemotherapy. I would like to nominate an old discovery of Dr. Liu published in 2008 in Biology Direct as the Breakthrough of this Year because, if one read some recent publications in top journals carefully, one should realize Lius theory or hypothesis makes more sense and shed more light on some enigmas in cancer research. For reference of Dr. Lius original discovery on this, please read: Occurrence of cancer at multiple sites: Towards distinguishing multigenesis from metastasis (Biology Direct 3: 14, 2008; http://www.biology-direct.com/content/3/1/14 ) 2010/12/13 , 11:44:42 Keming Cui Dear Science Editors, I am retired professor from Peking University, China. Who is Adam? Be divided into mainstream science and sidestream science what oneself is absurd. In those early years Mendel and Darwin once were the sidestream scientists in those early years science as well, but the history prove that they are great real scientists. If you if want to listen to you love to listen to, presses Adam to say of do, chase our opinion deletions. 2010/12/13 , 9:08:43 標記 Adam Breakthrough of the Year: Is mainstream science being overflowed by sidestream science? Dear Science Editors, It is amazing that several comments appeared in this serious website have might have played an amusing Liu symphony. Please check if all these different comments were sent by one man. If that was true, then there might be a spammer going around. If that is not true, then this may reflect a situation that the dam protecting mainstream science is being eroded by sidestream science. What should Science do now? Block this erosion? Or Let the sidestream science overrun the mainstream science? I think Science is at a very critical and also dangerous moment. If the science running in the sidestream contains more truth than that pushed in the mainstream, blocking the break-in of this sidestream in front of millions of web watchers would be a high-risk action. But let the sidestream flood the mainstream, then a breakthrough in science is really happening before our eyes. 2010/12/13 , 5:14:47 popo Dear Science Editors, I am an undergraduate student in China and got to know Dr. Shi Liu two years ago via his blog articles on iPS reprogramming at a very popular stem cell communication website in China ( http://www.stemcell8.cn/forum.php ). I found his criticisms on some flawed research in iPS reprogramming interesting. By digging into his publications, I found that his capability of seeing through the hype in iPS research is well based on his solid discoveries on cell life including stem cells ( http://im1.biz/StemCell.htm ). Also, his idea on cell divisions is quite interesting and insightful. At least it really changed the way I think of cell life and also lead me to reach a balanced view on iPS cells. We may not agree with all of his ideas, but we will defend to death his right to express his ideas. In the world of science, the solely rule is truth rather than fame and position. Nobody is always right, even Newton and Eistein had mistakes and bad ideas. iPS is a newly emerging field, at present, it is difficult to gauge how it will develop in the future. Thus, criticism from the minority like Dr. Liu shuld be respected. Li 2010-12-12 , 19:19:11 Ray ZHANG direct transdifferentiation in vivo, hoping the future stem cell therapy will not transplant the foreign stem cell... 今天 , 7:14:21 Best Biology I am a layman to biology. However, by reading many articles in LIU shi ( 刘实 )s very popular blog in Chinese ( http://blog.sina.com.cn/im1 ), I am convinced that his key discovery on cell life cell cannnot be divided but is reproduced is the most significant discovery in life science. However, that discovery may not be suitable for the honor of The Breakthrough of the Year becauase is is a breakthrough of centuries. Just think about how far we need to go back to correct the textbooks in biology! 2010-12-11 , 2:20:51 Science from China Just found: Science in China leading Science on bacterial aging research Dear Editors of Science: Several comments (seem very solid) have introduced Dr. Lius discoveries on iPS cells and iPS reprogramming and wished to convince people to accept a claim that it is Dr. Luis original discoveries that have changed the course of iPS research. I am not sure other people in the field would be willing to accept this and thus ask Science to be careful in reaching a decision. At a minimum, Science editors should read Dr. Lius publications and compare them with other publications to see if the later discoveries were truly some repeated discoveries (with more data) of Dr. Liu original discoveries (based on his analysis of previous published but limited data). More importantly, I urge editors to use a correct definition on discovery and respect insightful discoveries made without great cost and even before the availability of the rich data. Please think of the discovery of double helix for DNA and the importance of theoretical discoveries in science. However, on the studying of bacterial aging, I found that Dr. Liu is indeed a true pioneer. This is because I have just found that his much insightful discovery on this topis was published in 1999 in Science in China a peer-reviewed top journal in China and his publication appeared in both the English and the Chinese editions of the journal. Thus, his discovery in this field is even years earlier than the so-called first discovery published in Science in 2003 and PloS Biology in 2005. I also leant that Dr. filed a patent in 2000 based on his discovery and that patent was granted in 2004 by the US PTO. How could the whole western world miss such a very important discovery published in China?! I am wondering and outraged. See references: Liu, S. V. 1999. Tracking bacterial growth in liquid media and a new bacterial life model. Science in China (Series C: Life Science) (Chinese) 29:571-579. Liu, S. V. 1999. Tracking bacterial growth in liquid media and a new bacterial life model. Science in China (Series C: Life Science) (English) 42:644-654. Liu, S. V. 2004. Method and apparatus for producing age-synchronized cells. US patent US6767734B Ackermann, M., S. C. Stearns, and U. Jenal. 2003. Senescence in a bacterium with asymmetric division. Science 300:1920 Stewart, E. J., R. Madden, G. Paul, and F. Taddei. 2005. Aging and death in an organism that reproduces by morphologically symmetric division. PLoS Biol 3:295-300 訪客 The turning point of iPS research From beginning, iPSCs have been regarded as induced pluripotent stem cells that are indistinguishable from embryonic stem cells (ESCs) and thus can be ethically and safely utilized for regenerative medicine. However, an alternative view has been expressed in publications (see titles listed at http://im1.biz/iPS.htm ) that iPSCs are incorrectly programmed stem cells and thus can be distinguished from ESCs. It was further pointed out that iPSCs are man-made cancer stem cells (mm-CSCs) and can cause cancer upon transplantation. Now distinctions between iPSCs and ESCs are found in many aspects. The cancer risk of iPSCs has also been increasingly recognized. A very recent publication in Cell Stem Cell (7: 521-531, 2010) demonstrated the chromosomal aberrations in human iPSCs that are present in early passages and thus likely originated from the parent cells. Thus, iPS research may finally come to a turning point. Instead of being treated as the fountain of youth for regenerative medicine, iPSCs may serve as models for cancer research. Let us learn from what Yamanaka did in Science on August 1, 2008. He publically thanked Shi V. Liu (see Science 321:641, 2008; http://www.sciencemag.org/content/321/5889/641.full ) when he learnt some criticisms (rejected by Science but published in Logical Biol.8: 57-61, 2008). He even went on reporting cancer risk of his own iPS cells (see for examples, Nature Biotechnol. 27: 743-745, 2009 and PNAS 107: 14152-14157, 2010). Let us turn cancerous iPSCs for some really good use. 2010-12-10 , 15:32:29 Keming Cui Reprogramming out of the iPS detour I am a botanist, but watching closely the development of iPS and cell reprogramming research I am very happy to see that, after spending four years in a huge detour in the iPS track, cell reprogramming is now back to some straight ways. Looking retrospectively, I think we all should thank one man: Shi V. Liu. This is because he is the only single person who has fight very hard against a huge hype in iPS research and made all the discoveries on the true nature of iPS cells and iPS reprogramming. All the later experimental observations at most serve as some kinds of confirmations of his insightful pioneering discoveries. To verify this, one just needs to click this link http://im1.biz/iPS.htm and reads articles published by Shi V. Liu and then compares his publications from Truthfinding Cyberpress with later publications in the top journals. From a botanic point of view, dedifferentiation, redifferentiation and transdifferentiaon are some normal ways of plant life. But reprogramming a differentiated cell all way back to undifferentiated state and then differentiated to a desired cell is not only a waste of energy but also an opportunity for introducing abnormality. Our past experience in studying plant has taught us how easy it is to induce a normal cell via dedifferentiation into callus which is similar to the cancer cell in animals. Therefore, I fully understand Shi V. Liu 's views and found it is hard to believe that his very correct views have been intentionally neglected by animal cloners and cell reprogrammers. History does repeat sometimes. I remembered the Breakthrough of the Year in 2006 was The Poincar ConjectureProved (Science 314: 1848-1849, 2006). But that breakthrough research was published four years earlier in a non-peer-reviewed web archive called arXiv which even has no impact factor. 2010-12-10 , 11:42:36 標記 Responsible Science The real Breakthrough of the Year is the significantly increased retractions by CNS which stands for Correcting Non-sense in Science. 2010-12-10 , 10:59:50 訪客 Solving the structure of eukariotic ribosome is a pretty major discovery. 2010-12-10 , 0:42:22 訪客 Tumors grow their own blood vessels. 2010-12-09 , 10:00:55 訪客 arsenic life 2010-12-09 , 5:48:31 webber Dennis Lo: detecting fetal DNA in maternal blood 2010-12-08 , 21:19:30 kos Nyerges Synthetic genome controlled cell by JCVI 2010-12-08 , 19:07:29 訪客 The discovery of the enzyme Tet1 for the synthesis of 5 methyl-hydroxy cytosine by Aravind, Anjana Roa and colleagues (published in Science) is likely to open up new possibilities by acting as the sixth base in addition to A, T, G, C and meC. I think this is a fundamental discovery! Conversion of 5-methylcytosine to 5-hydroxymethylcytosine in mammalian DNA by MLL partner TET1. Impaired hydroxylation of 5-methylcytosine in myeloid cancers with mutant TET2. 2010-12-08 , 15:02:10 訪客 I don't know - something about extrasolar planets ? 2010-12-08 , 4:42:12 訪客 Quantum Physics Breakthrough: Scientists Find an Equation for Materials Innovation ScienceDaily (Feb. 26, 2010) Princeton engineers have made a breakthrough in an 80-year-old quandary in quantum physics, paving the way for the development of new materials that could make electronic devices smaller and cars more energy efficient. 2010-12-08 , 4:40:42 訪客 Quantum Entanglement Achieved in Solid-State Circuitry - Jan 12, 2010 2010-12-08 , 4:39:42 Kim Dudzik Oxygen on Rhea? Totally amazing. My personal pick for breakthrough of 2010. 2010-12-07 , 12:32:26 Rafael Roesler 1) Exposure to an enriched environment reduces cancer growth through a BDNF-dependent mechanism (Cao et al. http://www.cell.com/abstract/S0092-8674(10)00565-9) 2) Brain tumor stem cells give rise to endothelium (Wang et al., http://www.nature.com/nature/journal/vaop/ncurrent/full/nature09624.html) 2010-12-07 , 6:37:22 訪客 Last week's anouncement of aging reversal in the mouse 2010-12-06 , 9:30:00 說讚的人 訪客 Kevin It has to be the ability to look at a persons genes and figure out if they will live a long life. I can't wait to find out how long I will live! 2010-12-06 , 4:50:25 說讚的人 訪客 Ankur Sharma No doubt Synthetic life by Craig Venter 2010-12-06
干细胞的往日情怀 iPS still sings the way we were 细胞有记忆吗？ 作为一个完整的生物个体，记忆是其应对周围环境、维持生存的所必要的基本功能。对高等生物而言，记忆学习的基础。那么作为生物体基本构成单元的细胞是否也有记忆呢？答案是肯定的，但是，此记忆非彼记忆也。对于复杂的多细胞生物或高等生物而言，记忆常由特化的专门组织或器官来完成，如神经系统，其过程极其复杂，不是这里要讨论的。细胞的记忆则是由自己独立完成，它主要通过给基因挂上各种形式的记忆标签来实现的，这些标签又被称为表观遗传学信息。 我们知道，对于有性生殖的多细胞生物，机体的每一个不同的细胞都是来源于一个共同的祖先 - 受精卵，受精卵经过分裂和分化，形成各种组织细胞，最终组合成一个完整的个体。受精后 3-5 天形成的胚胎（又称囊胚）囊内的每一个细胞都可以单独发育成一个完整的个体，这就叫发育的全能型，或分化的全能型。能分化形成新的组织细胞类型的细胞，就是干细胞。当然，能够自我更新也是称之为干细胞一个必要的条件。干细胞是生命的原始种子细胞，在体内可以形成为各种组织细胞甚至器官和完整的生命体，因此具有潜在巨大的生物医学价值。 经过漫长和复杂分化历程形成的各种组织细胞的命运最终被定格：或是肌肉细胞、或是皮肤细胞等，细胞牢牢地记住了自己的分化经历，并终生严格维持自己最后的身份，而这些记忆是以表观遗传学信息形式储存。命运似乎不能任意改变的，这就是人们一直认为的分化过程是不可逆（而在植物，分化过程是可逆的，因此可以将植物任何部位的组织切下一块形成完整的植株）。这一观念直到 1997 年克隆羊多莉诞生才被彻底扭转。克隆多莉羊的技术被称为核移植技术，就是从供体羊（多莉的父亲或母亲）的身体取出单个体细胞，将细胞核吸出，移植到去除细胞核的卵子，让体细胞的细胞核载在卵子的细胞浆中逐渐忘掉分化过程所形成的成长记忆，将记忆回零以便重新开始，又叫重新编程。鉴于细胞核移植还涉及卵子的来源和使用，也一定程度存在伦理因素的制约。 于此同时，科学家们开始尝试用分子生物学方法直接对体细胞进行重新编程，使其回归到胚胎干细胞的状态。这在一开始被看做是不可能实现的任务（ impossible mission ），日本的山中伸弥实验室是最早啃这块硬骨头的人。 ２００７年，日本的山中伸弥实验室和随后美国 Thomson 实验室分别宣布通过组合特定四种控制细胞增殖和分化的基因人为表达实现将体细胞 ( 普通皮肤细胞 ) 成功地转化为干细胞的方法，被称为诱导多功能干细胞，又名 iPS (Induced pluripotent stem cell) 细胞。 iPS 技术绕开了胚胎干细胞研究一直面临的伦理和法律等诸多障碍，因此在医疗领域的应用前景非常广阔，于是 iPS 研究一时风起云涌，成为生物医学研究的最前沿热点，诱导技术在不断发展和成熟，先有基因转染，到后来的蛋白因子鸡尾酒，再后来又有华裔科学家丁盛的化学小分子组合技术等等。 仅管各个实验都声称有通过各种方法诱导获得的干细胞和胚胎干细胞完全一样，但是人们仍然会心存疑虑：体细胞的原有记忆是否真正被完全抹去，可以完全重新开始？因为残存的记忆会引导或干扰干细胞的分化行为，恰如一个有着长久偷窃行为记忆的人可能不自觉地会对别人的口袋感兴趣；而一个有着毒品成瘾记忆的人可能在绝望或沮丧时可能更容易想到毒品。在 7 月 19 日 Nature 和其姊妹期刊 Nature Biochenology 分别发表了来自美国多个不同实验室的研究结果，证实了这个担忧并非杞人忧天。 首先，由哈佛的学和约翰霍普金斯大学的两个著名实验室合作，比较了早期 iPS 和体细胞核移植获得的干细胞在表观遗传标记上的变化，如 DNA 上的甲基化，结果发现，早期的 iPS 细胞仍然残存一定数量的甲基化，说明 iPS 憨部分保留着原有的记忆，而这些表观遗传学记忆必定影响细胞的分化过程中的行为。 另外发表在 Nature Biochenology 的是由多大六个实验室的合作研究的结果。该研究比较了不同组织来源的体细胞在经过标准的诱导方法获得 iPS 后在分子水平和分化行为上的差异。细胞的特征和功能取决于其多种基因的表达状态（表达或不表达，或表达水平），而这些基因的表达状态就是细胞的基因表达谱，相当于细胞的脸谱，基因芯片技术是描述细胞脸谱的一个主要技术。该研究发现：不同来源的细胞制成的 iPS 基因表达谱差异很大，说明这些细胞还都保存着过去的记忆。将这些不同细胞来源的 iPS 在培养瓶内进行诱导分化实验，结果发现， iPS 倾向于分化成它们原有的、或谱系接近的细胞类型，例如，从造血细胞来源的细胞诱导形成的 iPS 在培养瓶内很容易就可以诱导分化成造血来源的细胞，但很难形成神经系统的细胞。也就是说这些残存的记忆更容易引导这些细胞想到或回到过去的往日情怀（ The Way We Were ）。 iPS 的往日情怀是阻碍其多向分化的障碍，这两篇报道着也实让我们虚惊了一场。所幸的是，这种记忆也并非多么顽固。时间可以淡化或抹去记忆：如果把 iPS 在培养瓶中再继续培养几星期，或者几个月，那么他们原来的记忆印迹就将逐渐消失，恰如芭芭拉 . 史翠珊所唱歌词中期望的：让时间重写记忆，让一切重新开始。 iPS 的前景依然是光明的，虽然还有很长路要走，而干细胞向实际应用可能还有更长的路要走。期待着这么一天，成熟的干细胞技术的能帮助那些饱受病痛折磨的人们驱走梦魇，重新开始。 文章链接 http://www.nature.com/nature/journal/vnfv/ncurrent/full/nature09342.html （ http://www.nature.com/nbt/journal/vaop/ncurrent/abs/nbt.1667.html ） The Way we were memories light the corners of my mind misty water color memories of the way we were scattered pictures of the smiles we left behind smiles we gave to one another fore the way we were can't it be that it was all so simple then or has time rewritten every line and if we had the chance to do it all again tell me would we, could we memories maybe beautiful and yet what's too painful to remember we simply choose to forget so it's the laughter, we will remember whenever we remember the way we were
标签: iPS
