STEM CELL PRODUCTION CORE

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。 间充质干细胞或骨髓基质细胞(MSCs)是来自骨髓的成体干细胞的一个子集。这些细胞具有医学和治疗价值，因为它们已被证明可分化为成骨细胞、脂肪细胞、软骨细胞、肌细胞、星形胶质细胞、少突胶质细胞和神经元。由于其固有的可塑性，这些细胞有可能用于治疗大量的遗传性疾病。Bunnell博士已经成功地确定了恒河猴骨髓间充质干细胞的扩增和特性要求。干细胞生产核心设施(SCPC)专注于非人类灵长类MSCs的产生、维持和分布。我们常规从恒河猴骨髓和脂肪组织标本中制备间充质干细胞。目前，我们有一个从100多只不同年龄的恒河猴中产生的骨髓间充质干细胞细胞系的库，这些细胞系已经准备好并准备分发。SCPC目前不仅对基因治疗计划有很大的影响，而且对TNPRC内的其他部门，如TNPRC内的比较病理学，以及杜兰健康科学中心和彭宁顿生物医学研究中心内的部门和中心以及全国其他研究实验室都有很大影响。 为合作研究项目生产恒河猴干细胞制剂。为此，我们计划使用在上一个资助期开发的技术来分离干细胞(MSCs、NSPC和ESCs)，以扩大从不同年龄(胎儿、新生儿和成人)和不同性别的动物分离的各种干细胞库。这些细胞将被用来生成大型细胞库，供合作科学家在内部或外部使用。此外，我们将与有兴趣进行自体移植研究的科学家合作，为他们的研究项目生成特定动物的MSC库。 继续描述非人灵长类干细胞的特性。我们在这些研究中的目标是描绘不同干细胞群体在成熟和/或分化过程中发生的细胞和分子事件。该部门的科学家将继续在体外比较各种干细胞的生物学特性。将产生能够最大限度地促进MSCs及其后代分化和增殖的恒河猴干细胞特异性体外分化的检测方法。体外鉴定的一个焦点将是所描述的所有干细胞沿神经谱系的分化。为了完成克拉贝氏病的治疗，干细胞需要分化为少突胶质细胞。为实现这一目标，将作出重大努力，开发和应用差异化协议。 全面开发检测干细胞植入的检测方法。最重要的是，该部门将在评估非人类灵长类干细胞在小鼠和非人类灵长类动物模型中体内植入的分析方法的产生中发挥重要作用。在过去的资助期间，该部门建立了使用人类核抗原抗体在小鼠模型中常规检测NHP细胞的技术，这很容易检测到NHP细胞。该部门对BrdU进行了适当的染色，并对EGFP进行了直接免疫荧光评估。在下一个资助期，该部门将需要专注于检测NHP组织中的NHP细胞，使用Y染色体原位杂交或使用单核苷酸多态(SNPs)检测细胞。这将使我们有机会与兽医司的遗传学和基因组库核心以及细菌学和寄生虫学的DNA微阵列和表达核心就单核苷酸多态性在干细胞检测中的应用进行合作，因为这些核心将开发用于亲本血统测试的SNP技术。 结合干细胞和基因转移技术用于恒河猴Krabbe病模型。我们的目标是在体外和体内确定干细胞及其分化后代对有效基因传递和表达的要求。我们将对我们的各种非人类灵长类干细胞载体系统的基因转移效率进行表征。这将使我们有机会产生标记有报告基因元件、绿色荧光蛋白或β-半乳糖苷酶的干细胞库。更重要的是，我们将评估为恒河猴骨髓间充质干细胞移植后在体外和体内提供高水平和长期基因表达的要求。

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. Mesenchymal stem cells or marrow stromal cells (MSCs) are a subset of adult stem cells from bone marrow. These cells are of medical and therapeutic interest because they have been shown to differentiate into osteoblasts, adipocytes, chondrocytes, myocytes, astrocytes, oligodendrocytes and neurons. Due to their inherent plasticity, these cells have the potential to be useful for the treatment of a large number of genetic diseases. Dr. Bunnell has successfully defined the requirements for the expansion and characterization of rhesus macaque MSCs. The Stem Cell Production Core Facility (SCPC) focuses on generation, maintenance and distribution of nonhuman primate MSCs. We routinely prepare MSCs from rhesus macaque bone marrow and adipose tissue samples. We presently have a bank pf MSC cell lines generated from over 100 rhesus macaques of varying age prepared and ready for distribution. The SCPC presently has a large impact not only on the Gene Therapy Program, but on other divisions such as Comparative Pathology within the TNPRC, but also Departments and Centers within the Tulane Health Sciences Center and the Pennington Biomedical Research Center and other research labs nationally. Produce rhesus macaque stem cell preparations for collaborative research projects. For this we plan to use the techniques developed over the last funding period to isolate stem cells (MSCs, NSPCs and ESCs) to expand the bank of various stem cell types isolated from animals of various ages (fetal, newborn and adults) and sexes of animals. The cells will be used to generate large cell banks for use internally or externally by collaborating scientists. Additionally, we will work with scientists interested in performing autologous transplantation studies by generating MSC banks from specific animals for their research programs. Continue the characterization of nonhuman primate stem cells. Our goal in these studies will be to delineate the cellular and molecular events that take place during maturation and/or differentiation of the various stem cell populations. Scientists in the division will continue to compare the biologic properties of the various stem cells in vitro. Assays mediating in vitro differentiation specific to rhesus stem cells that will maximize the differentiation and proliferation of MSCs and their progeny will be generated. One focus of the in vitro characterization will be the differentiation along neural lineages for all of the stem cells described. In order to accomplish the treatment of Krabbe's disease, stem cells will need to undergo differentiation into oligodendrocytes. A significant effort will be made towards the development and application of differentiation protocols to accomplish this goal. Fully develop assays for the detection of the engraftment of stem cells. Most importantly, the division will play a significant role in the generation of assays for the assessment of engraftment of nonhuman primate stem cells in vivo in both murine and nonhuman primate models. Over the last funding period, the division established technologies for the routine detection of NHP cells in murine models using antibodies to human nuclear antigen, which readily permits the detection of NHP cells. The division has in place staining for BrdU and direct immunofluorescence assessment of EGFP. Over the next funding period, the division will need to focus on the detection of NHP cells in NHP tissues, using technologies such as in situ hybridization for the Y chromosome or detection of cells using single nucleotide polymorphisms (SNPs). This will provide us an opportunity to collaborate with the Genetics and Genome Banking core in the Division of Veterinary Medicine and the DNA Microarray and Expression Core in Bacteriology and Parasitology on the application of SNPs for stem cells detection, as these cores will be developing SNP technologies for the testing of parental lineages. Combining stem cell and gene transfer technologies for use in the rhesus Krabbe's disease model. Our goal is to characterize the requirements for effective gene delivery and expression in stem cells and their differentiated progeny both in vitro and in vivo. The gene transfer efficiency of our various vector systems for nonhuman primate stem cells will be characterized. This will permit us the opportunity to generate banks of stem cells labeled with reporter gene elements, EGFP or beta-galactosidase. More importantly, we will assess the requirements that provide for high levels and protracted duration of gene expression in rhesus MSCs both in vitro and in vivo following transplantation.