WNPRC STEM CELL RESOURCE

该子项目是利用 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH来源获得主要资金， 因此可以在其他CRISP条目中表示。列出的机构是 中心，不一定是研究者的机构。 目的：维持和进一步发展与多能干细胞研究有关的专门研究资源。 资源访问分配： 到目前为止，威斯康星州国家灵长类动物研究中心的干细胞资源单位向感兴趣的研究人员提供冷冻的恒河猴和绒猴ES细胞。没有拒绝任何请求。此外，干细胞资源单元提供斑马鱼bFGF用于培养灵长类动物ES细胞。自上次提交以来，3名新研究者接受了重组蛋白，使定期接受zbFGF的研究者总数达到21名。迄今为止，已分发了超过330 mg的zbFGF。这为调查人员节省了100多万美元。用于纯化蛋白质本身的DNA质粒现在可以通过Addgene（www.addgene.com）获得，并于2008年发送给13名新的研究人员，使接受该质粒的研究人员总数达到39人。最后，在2009年，2名研究者要求并获得了由干细胞资源部门提供并由WiCell研究所分发的恒河猴ES细胞。 传播： 通过在同行评审期刊上发表文章和出席科学会议，向科学界传播知识。威斯康星州国家灵长类动物研究中心还举行季度研究务虚会，以增加各种服务和资源单位之间的沟通。 培训内容： 灵长类动物胚胎干细胞培养技术的培训是可用的。许多新调查员在以往报告所述期间利用了这一资源，但今年没有新调查员接受培训。 进度： 干细胞资源的过去和现在的成员开发了一种在没有载体转基因序列的情况下产生iPS细胞的方法;这项工作在下面的重点部分中引用。我们已经为几个UW研究者提供了疾病特异性细胞系的ips细胞衍生。到目前为止，已经有4种疾病细胞系被用于诱导多能干细胞的产生，总共有55个克隆被培养和冷冻以备将来使用。我们也开始在恒河猴成纤维细胞上进行重编程实验。其他研究小组已经成功地对灵长类细胞进行了重编程，我们将遵循类似的协议。 亮点： 干细胞资源的过去和现在的成员是一篇论文的作者，该论文详细介绍了成人细胞重编程的无载体方法：人类诱导多能干细胞无载体和转基因序列Junying Yu，Kejin Hu，Kim Smuga-Otto，Shulan Tian，罗恩斯图尔特，Igor I。作者声明：A.汤姆森科学2009年5月8日2009年10月324日。第5928号，第797-801页。 挑战： 由于资金短缺，我们无法接收食蟹猴胚胎，该项目被迫停止。我们期待着与CPI合作，接收食蟹猴胚胎，以创建新的食蟹猴ES细胞系，如我们的P51提案中所述。 关注事项： 目前没有问题。 干细胞资源支持涉及许多部分或全部依赖于WNPRC资源的期刊文章。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Objective: To maintain and further develop a specialized resource for studies relating to pluripotent stem cell research. Allocation of Resource Access: To date, the stem cell resource unit at the Wisconsin National Primate Research Center provides frozen rhesus and marmoset ES cells to interested investigators. No request has been denied. Additionally, the stem cell resource unit provides zebrafish bFGF for culturing primate ES cells. Since last submission 3 new investigators have received the recombinant protein bringing the total number of investigators receiving zbFGF on a regular basis to 21. Over 330mg of zbFGF has been distributed to date. This has saved investigators over $1,000,000. The DNA plasmid used to purify the protein itself is now available through Addgene (www.addgene.com) and was sent to 13 new investigators in 2008 bringing the total number of investigators to receive this plasmid to 39. Lastly, in 2009, 2 investigators requested and received rhesus ES cell provided by the stem cell resources unit and distributed by the WiCell Research Institute. Dissemination: Knowledge is disseminated to the scientific community via publications in peer reviewed journals and scientific meeting attendance. The Wisconsin National Primate Research Center also holds quarterly research retreats to create increased communication between the various service and resource units. Training: Training in culture techniques of primate embryonic stem cells is available. Many new investigators have taken advantage of this resource in previous reporting periods however there have been no new investigators trained this year. Progress: Past and present members of stem cell resources developed a method for generating iPS cells without vector transgene sequences; this work is referenced below in the highlights portion. We have provided ips cell derivation for disease specific cell lines for several UW investigators. To date, 4 disease cell lines have been used for ips cell generation and a total of 55 clones have been cultured and frozen for future use. We are also beginning to start reprogramming experiments on rhesus macaque fibroblasts. Other groups have successfully reprogrammed primate cells and we will be following similar protocols. Highlights: Past and present members of stem cell resources were authors on a paper detailing a vector free method of adult cell reprogramming: Human Induced Pluripotent Stem Cells Free of Vector and Transgene Sequences Junying Yu, Kejin Hu, Kim Smuga-Otto, Shulan Tian, Ron Stewart, Igor I. Slukvin, James A. Thomson Science 8 May 2009 Vol 324. No. 5928, pp 797-801. Challenges: Due to funding shortages we were unable to receive cynomologous embryos and the project came to a halt. We look forward to working with CPI to receive Mauritian cynomolougous embryos to create new cynomologous ES cell lines as described in our P51 proposal. Concerns: No concerns at this time. Stem Cell Resource support is involved in numerous journal articles that depend in part or in full on WNPRC resources.