Cell Stem Cell揭示受精卵DNA去甲基化重要机制

<正>2014年9月11日,Cell Stem Cell在线发表了中国科学院上海生科院生物化学与细胞生物学研究所徐国良研究组、李劲松研究组和北京大学生命科学学院生物动态光学成像中心汤富酬研究组的最新研究成果,该研究发现小鼠早期胚胎中母源和父源基因组在单细胞的受精卵阶段均会发生大规模的DNA主动和被动去甲基化,并且DNA双加氧酶Tet3介导了主动去甲基化的发生,而糖苷酶     

A Special Issue on Stem Cell Research

Stem cells possess the remarkable ability of extensive self-renewal and differentiation into specific celllineages,and they play essential roles in development and adult tissue homeostasis.Due to their criticalimportance in normal physiology and the promise for use in regenerative medicine to treat a variety ofdiseases,stem cells have attracted extensive research interest in recent years.As China's premium in-ternational journal with a broad scope in cell and molecular biology,Cell Research has witnessed moreand more submissions on stem cell research.Indeed,along with the traditional strengths of the journal inmolecular immunology,cancer biology,and plant molecular physiology,stem cell research has graduallyand naturally evolved into a new growth point of Cell Research.Reflecting the growing interest of bothour readers and authors in this exciting and expanding field,we are pleased to present this Special Issueon Stem Cell Research.     

Cell Stem Cell发表原始多潜能干细胞的最新研究成果

<正>2014年10月2日,Cell Stem Cell刊登了北京大学生命科学学院邓宏魁研究组最新研究成果。该研究在世界上首次成功将恒河猴皮肤细胞诱导成为了"原始多潜能干细胞",是干细胞领域的一项重要突破。北京大学生命科学学院邓宏魁研究团队早在2009年在国际上首次建立传统的恒河猴诱导多能干细胞(iPS细胞),在这个研究基础上,他们发现一些关键信号通路的调控因子,将恒河猴的传统     

Is Ewing's Sarcoma a Stem Cell Tumor?

Solving the histogenesis of Ewing's sarcoma has defied investigators despite progress in understanding its molecular pathogenesis. In a recent issue of Cancer Cell, Tirode et al. (2007) present evidence supporting the hypothesis that this rare cancer arises from a primitive mesenchymal precursor.     

Cancer: Inappropriate Expression of Stem Cell Programs?

Cancer stem cells (CSCs) are a subpopulation of cancer cells that possess characteristics, including self-renewal, associated with normal stem cells. In this issue of Cell Stem Cell, Wong et al. (2008) define a core embryonic stem cell (ESC)-like gene expression program that may be important for CSC function in multiple epithelial cancers.     

Quantification of Biomass and Cell Motion in Human Pluripotent Stem?Cell?Colonies

Somatic cell reprogramming to pluripotency requires an immediate increase in cell proliferation and reduction in cell size. It is unknown whether proliferation and biomass controls are similarly coordinated with early events during the differentiation of pluripotent stem cells (PSCs). This impasse exists because PSCs grow in tight clusters or colonies, precluding most quantifying approaches. Here, we investigate live cell interferometry as an approach to quantify the biomass and growth of HSF1 human PSC colonies before and during retinoic acid-induced differentiation. We also provide an approach for measuring the rate and coordination of intracolony mass redistribution in HSF1 clusters using live cell interferometry images. We show that HSF1 cells grow at a consistent, exponential rate regardless of colony size and display coordinated intracolony movement that ceases with the onset of differentiation. By contrast, growth and proliferation rates show a decrease of only ∼15% decrease during early differentiation despite global changes in gene expression and previously reported changes in energy metabolism. Overall, these results suggest that cell biomass and proliferation are regulated independent of pluripotency during early differentiation, which is distinct from what occurs with successful reprogramming.     

Glioma Cell Death: Cell–Cell Interactions and Signalling Networks

The prognosis for patients with malignant gliomas is poor, but improvements may emerge from a better understanding of the pathophysiology of glioma signalling. Recent therapeutic developments have implicated lipid signalling in glioma cell death. Stress signalling in glioma cell death involves mitochondria and endoplasmic reticulum. Lipid mediators also signal via extrinsic pathways in glioma cell proliferation, migration and interaction with endothelial and microglial cells. Glioma cell death and tumour regression have been reported using polyunsaturated fatty acids in animal models, human ex vivo explants, glioma cell preparations and in clinical case reports involving intratumoral infusion. Cell death signalling was associated with generation of reactive oxygen intermediates and mitochondrial and other signalling pathways. In this review, evidence for mitochondrial responses to stress signals, including polyunsaturated fatty acids, peroxidising agents and calcium is presented. Additionally, evidence for interaction of glioma cells with primary brain endothelial cells is described, modulating human glioma peroxidative signalling. Glioma responses to potential therapeutic agents should be analysed in systems reflecting tumour connectivity and CNS structural and functional integrity. Future insights may also be derived from studies of signalling in glioma-derived tumour stem cells.     

Cell Stem Cell:线粒体DNA突变潜藏于人诱导性多能干细胞中

正在一项新的研究中,美国俄勒冈健康与科学大学(OHSU)胚胎细胞与基因治疗中心主任Shoukhrat Mitalipov博士及其研究团队首次证实一个存在已久的假设:当变老时,基因突变积累在人们的线粒体(即细胞能量工厂)中。研究人员发现作为一类对病人的皮肤细胞或血细胞进行重编程而产生的干细胞,诱导性多能干细胞(iPS细胞)含有存在缺陷的线粒体DNA。相关研究结果在     

Proliferating Patent Problems with Human Embryonic Stem Cell Research?

The scientific challenges and ethical controversies facing human embryonic stem cell (hESC) research continue to command attention. The issues posed by patenting hESC technologies have, however, largely failed to penetrate the discourse, much less result in political action. This paper examines U.S. and European patent systems, illustrating discrepancies in the patentability of hESC technologies and identifying potential negative consequences associated with efforts to make available hESC research tools for basic research purposes while at same time strengthening the position of certain patent-holders' rights. Differences between the U.S. and the European contexts may in part explain why the course of hESC research in those jurisdictions ultimately diverges. Nevertheless, questions about whether and how patenting, related agreements, and licensing practices progress and shape the field of hESC research in both the U.S., Europe, and elsewhere must no longer be marginalised. These questions are fundamentally important in determining what benefits are likely to result from hESC research. Assuring these benefits is the moral issue with which patent systems are most intrinsically concerned, and that governments must begin to directly address rather than assume or ignore.     

Cell Stem Cell:线粒体DNA突变潜藏于人诱导性多能干细胞中

正在一项新的研究中,美国俄勒冈健康与科学大学(OHSU)胚胎细胞与基因治疗中心主任Shoukhrat Mitalipov博士及其研究团队首次证实一个存在已久的假设:当变老时,基因突变积累在人们的线粒体(即细胞能量工厂)中。研究人员发现作为一类对病人的皮肤细胞或血细胞进行重编程而产生的干细胞,诱导性多能干细胞(iPS细胞)含有存在缺陷的线粒体DNA。相关研究结果在线发表在Cell Stem Cell期刊上。     

Meeting Announcement 2007 Shanghai International Symposium on Stem Cell Research

We are delighted to announce an international symposium on stem cell research to be held in Shanghai on November 6-9, 2007. This meeting is co-organized by Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences and several other research institutions in China, together with the International Society for Stem Cell Research （ISSCR）. This is the first time ISSCR co-organizes a meeting with other organizations. ISSCR will organize a delegacy of 8-10 members to participate the symposium.[第一段]     

Mass Spectrometry–based Proteomic Analysis of the Matrix Microenvironment in Pluripotent Stem Cell Culture

The cellular microenvironment comprises soluble factors, support cells, and components of the extracellular matrix (ECM) that combine to regulate cellular behavior. Pluripotent stem cells utilize interactions between support cells and soluble factors in the microenvironment to assist in the maintenance of self-renewal and the process of differentiation. However, the ECM also plays a significant role in shaping the behavior of human pluripotent stem cells, including embryonic stem cells (hESCs) and induced pluripotent stem cells. Moreover, it has recently been observed that deposited factors in a hESC-conditioned matrix have the potential to contribute to the reprogramming of metastatic melanoma cells. Therefore, the ECM component of the pluripotent stem cell microenvironment necessitates further analysis.In this study we first compared the self-renewal and differentiation properties of hESCs grown on Matrigel™ pre-conditioned by hESCs to those on unconditioned Matrigel™. We determined that culture on conditioned Matrigel™ prevents differentiation when supportive growth factors are removed from the culture medium. To investigate and identify factors potentially responsible for this beneficial effect, we performed a defined SILAC MS-based proteomics screen of hESC-conditioned Matrigel™. From this proteomics screen, we identified over 80 extracellular proteins in matrix conditioned by hESCs and induced pluripotent stem cells. These included matrix-associated factors that participate in key stem cell pluripotency regulatory pathways, such as Nodal/Activin and canonical Wnt signaling. This work represents the first investigation of stem-cell-derived matrices from human pluripotent stem cells using a defined SILAC MS-based proteomics approach.The two defining characteristics of human embryonic stem cells (hESCs),¹ self-renewal and pluripotency, are maintained by a delicate balance of intracellular and extracellular signaling processes. Extracellular regulation is primarily the result of changes in the microenvironment surrounding the cells during growth in vitro or in vivo. HESCs interact with this “niche ” through support cells, extracellular matrix (ECM) components, and autocrine/paracrine signaling (reviewed in Refs. 1–3). Modulation of any of these supportive elements individually or in combination has been used extensively to alter hESC behavior (1–3).The culture of hESCs, as well as that of human induced pluripotent stem cells (hiPSCs), is conventionally performed on a layer of irradiated mouse embryonic fibroblast cells (MEFs). These MEFs are believed to promote the maintenance of hESCs and hiPSCs through the secretion of beneficial support proteins and cytokines into the soluble microenvironment. A number of proteomic studies have been conducted that examine the secretome of feeder-cell layers in an attempt to elucidate proteins and pathways essential for hESC and hiPSC survival (4–7). Alternatively, hESCs and hiPSCs can be cultured in feeder-free conditions in the absence of support cells. In feeder-free conditions, hESCs and hiPSCs are most often grown on the basement membrane matrix Matrigel™ in medium that has been previously conditioned by MEFs (MEF-CM). Matrigel™ is a gelatinous mixture that is secreted by Engelbreth-Holm-Swarm mouse sarcoma cells (8). Although recent studies have proposed that a variety of defined matrices can support the growth of hESCs and hiPSCs, few of these can maintain a wide range of stem cell lines and therefore are typically not used in place of Matrigel™. The properties of Matrigel™ that make it such an effective matrix for hESC and hiPSC culture remain poorly understood. Because of the complexity of matrices like Matrigel™, the majority of proteomic studies that examine the hESC and hiPSC microenvironment have focused on contributions from support cells and soluble extracellular factors.The ECM is typically a complex network of structural proteins and glycosaminoglycans that function to support cells through the regulation of processes such as adhesion and growth factor signaling (9). Thus, it is not surprising that the generation of a well-defined matrix capable of facilitating hESC and hiPSC self-renewal has remained difficult (10). Previous proteomic investigations of Matrigel™ and other matrices supportive of hESC maintenance in vitro have revealed the presence of numerous growth, binding, and signaling proteins (11, 12). Further examination of how hESCs and hiPSCs interact with these complex matrices would provide critical information about what role the ECM plays in the organization of processes involved in the regulation of self-renewal and pluripotency.A recent study has established the ability of hESC-derived matrix microenvironments to alter tumorigenic properties through the reprogramming of metastatic melanoma cells (13). Importantly, this effect was found to be dependent on the exposure of metastatic cells to hESC-derived conditioned Matrigel™. Culture of metastatic melanoma cells in hESC-conditioned medium did not promote the reprogramming effect. These data suggest that the proteins responsible for this effect were integrated in the matrix. With the use of immunochemical techniques, it was later found that the left-right determination (Lefty) proteins A and B that were deposited in the matrix by hESCs during conditioning were at least in part responsible for the cellular change observed in metastatic cells (14). The Lefty A and B proteins are antagonists of transforming growth factor (TGF)-β signaling that act directly on Nodal protein, a critical regulator of the stem cell phenotype (15, 16). Subsequent studies of conditioned matrix utilizing mESCs implicated the bone morphogenic protein (BMP) 4 antagonist Gremlin as a primary regulator of the observed changes in metastatic cells (17). Collectively, these studies were all biased by a targeted analysis of potential effectors of metastatic cells. A comprehensive proteomic analysis of conditioned matrix could potentially reveal other factors involved in metastatic cell reprogramming. Furthermore, proteomic examination of hESC and hiPSC conditioned matrix could expose factors important in the regulation of self-renewal and pluripotency by the microenvironment in vitro.To this end, we have analyzed both types of human pluripotent stem cells, hESCs and hiPSCs, via a mass spectrometry (MS)-based proteomics approach to identify proteins deposited during growth in feeder-free conditions in vitro on Matrigel™. To investigate the hESC- and hiPSC-derived matrix, the metabolic labeling technique known as stable isotope labeling with amino acids in cell culture (SILAC) was used (18). SILAC facilitates the identification of hESC- and hiPSC-derived proteins that would otherwise be confounded by the presence of mouse-derived protein background from Matrigel™. From the proteomic analysis of three cells lines, namely, the hESC lines H9 and CA1 and the hiPSC line BJ-1D, we identified a total of 621, 1355, and 1350 total unique proteins, respectively. This work represents the first analysis of a hESC- and hiPSC-derived conditioned matrix and resulted in the identification of at least one novel microenvironmental contributor responsible for the regulation of human pluripotent stem cells.     

Cell Stem Cell:研究开发出在体外长期培养成体干细胞的方法

正在一项新的研究中,来自美国麻省总医院(MGH)等机构的研究人员开发出的一种新方法可能引发成体干细胞培养领域变革。研究人员描述了获得来自在日常治疗肺部疾病期间收集的各种组织样品中的气道干细胞(airway stem cell),并对它们进行增殖。这种方法似乎也可用于几种其他的组织,如皮肤、胃肠道内壁和生殖道。相关研究结果在线发表在Cell Stem Cell期刊上。     

Gap Junction–mediated Cell–Cell Communication Modulates Mouse Neural Crest Migration

Previous studies showed that conotruncal heart malformations can arise with the increase or decrease in α₁ connexin function in neural crest cells. To elucidate the possible basis for the quantitative requirement for α₁ connexin gap junctions in cardiac development, a neural crest outgrowth culture system was used to examine migration of neural crest cells derived from CMV43 transgenic embryos overexpressing α₁ connexins, and from α₁ connexin knockout (KO) mice and FC transgenic mice expressing a dominant-negative α₁ connexin fusion protein. These studies showed that the migration rate of cardiac neural crest was increased in the CMV43 embryos, but decreased in the FC transgenic and α₁ connexin KO embryos. Migration changes occurred in step with connexin gene or transgene dosage in the homozygous vs. hemizygous α₁ connexin KO and CMV43 embryos, respectively. Dye coupling analysis in neural crest cells in the outgrowth cultures and also in the living embryos showed an elevation of gap junction communication in the CMV43 transgenic mice, while a reduction was observed in the FC transgenic and α₁ connexin KO mice. Further analysis using oleamide to downregulate gap junction communication in nontransgenic outgrowth cultures showed that this independent method of reducing gap junction communication in cardiac crest cells also resulted in a reduction in the rate of crest migration. To determine the possible relevance of these findings to neural crest migration in vivo, a lacZ transgene was used to visualize the distribution of cardiac neural crest cells in the outflow tract. These studies showed more lacZ-positive cells in the outflow septum in the CMV43 transgenic mice, while a reduction was observed in the α₁ connexin KO mice. Surprisingly, this was accompanied by cell proliferation changes, not in the cardiac neural crest cells, but in the myocardium— an elevation in the CMV43 mice vs. a reduction in the α₁ connexin KO mice. The latter observation suggests that cardiac neural crest cells may have a role in modulating growth and development of non–neural crest– derived tissues. Overall, these findings suggest that gap junction communication mediated by α₁ connexins plays an important role in cardiac neural crest migration. Furthermore, they indicate that cardiac neural crest perturbation is the likely underlying cause for heart defects in mice with the gain or loss of α₁ connexin function.