WNPRC STEM CELL RESOURCE

这个子项目是许多研究子项目中利用 资源由NIH/NCRR资助的中心拨款提供。子项目和 调查员(PI)可能从NIH的另一个来源获得了主要资金， 并因此可以在其他清晰的条目中表示。列出的机构是 该中心不一定是调查人员的机构。 目的：维护和进一步发展与多能干细胞研究相关的专业资源。 资源访问权限分配： 到目前为止，威斯康星州国家灵长类研究中心的干细胞资源部向感兴趣的研究人员提供冷冻的恒河猴和绒猴ES细胞。没有任何申请被拒绝。此外，干细胞资源部还为培养灵长类胚胎干细胞提供斑马鱼碱性成纤维细胞生长因子。自上次提交以来，已有3000名新的调查人员收到了重组蛋白，使定期接受zbFGF的调查人员总数达到21人。到目前为止，已经分发了超过330毫克的zbFGF。这为调查人员节省了100多万美元。用于提纯蛋白质本身的DNA质粒现在可以通过Addgene(www.addgene.com)获得，并于2008年被发送给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，2009年，2名调查人员请求并收到了干细胞资源股提供、Wicell研究所分发的恒河猴胚胎干细胞。 传播： 通过在同行评议的期刊上发表出版物和出席科学会议，向科学界传播知识。威斯康星州国家灵长类研究中心还举行季度研究务虚会，以增加各服务和资源单位之间的沟通。 培训： 灵长类胚胎干细胞培养技术方面的培训是可用的。许多新的调查员在前几个报告期利用了这一资源，但今年没有新的调查员接受培训。 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；这项工作在下文的重点部分中被引用。我们已经为几个UW研究人员提供了疾病特异性细胞系的iPS细胞来源。到目前为止，已有4个疾病细胞系用于iPS细胞的生成，共培养并冷冻了55个克隆以供将来使用。我们还开始对恒河猴成纤维细胞进行重新编程实验。其他小组已经成功地对灵长类细胞进行了重新编程，我们将遵循类似的方案。 亮点： 过去和现在的干细胞资源成员都是一篇详细介绍成体细胞重新编程的无载体方法的论文的作者：人类诱导多能干细胞无载体和转基因序列于俊英，胡可进，Kim smuga-Otto，Shulan Tian，Ron Stewart，Igor I.Slukvin，James A.Thomson Science，2009年5月，第324卷。第5928号，第797-801页。 挑战： 由于资金短缺，我们无法获得细胞学胚胎，该项目陷入停顿。我们期待着与CPI合作，获得毛里求斯食蟹猴胚胎，以创建我们P51提案中描述的新的食蟹性胚胎干细胞系。 关注事项： 目前没有任何顾虑。 干细胞资源支持涉及许多期刊文章，这些文章部分或全部依赖WNPRC的资源。

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.