Yang Guang, Hong Hyenjong, Torres April, Malloy Kristen E, Roy-Choudhury Gourav, Kim Jeffrey, Daadi Marcel M
Southwest National Primate Research Center, Texas Biomedical Research Institute, San Antonio, TX, USA.
Department of Radiology, Research Imaging Institute, Cell Systems and Anatomy, Long School of Medicine, University of Texas Health Science Center at San Antonio, San Antonio, TX, USA.
Methods Mol Biol. 2019;1919:175-186. doi: 10.1007/978-1-4939-9007-8_13.
Limited access to primary tissue from various nonhuman primate (NHP) species represents a significant unmet need that hampers progress in understanding unique cellular diversity and gene regulation of specific tissues and organs in stem cell translational research. Most comparative biology studies have been limited to using postmortem tissue usually frozen specimens with limited utility for research. The generation of induced pluripotent stem cell (iPSC) lines from somatic cells, such as adult skin or blood cells, offers an alternative to invasive and ethically controversial interventions for acquiring tissue. Pluripotent iPSCs have virtually an unlimited capacity to proliferate and differentiate into all cell types of the body. We are generating high-quality validated NHP iPSC lines to offer to scientific community and facilitate their research programs. We use the non-integrative episomal vector system to generate iPSCs from NHP skin biopsies. In this chapter we describe the validation of NHP iPSC lines by confirming pluripotency and their propensity to differentiate into all three germ layers ectoderm, mesoderm, and endoderm according to established standards and measurable limits for a set of marker genes incorporated into a scorecard.
获取来自各种非人灵长类动物(NHP)物种的原代组织存在很大困难,这是一个尚未满足的重大需求,阻碍了干细胞转化研究中对特定组织和器官独特细胞多样性及基因调控的理解进展。大多数比较生物学研究仅限于使用死后组织,通常是冷冻标本,其研究效用有限。从体细胞(如成人皮肤或血细胞)生成诱导多能干细胞(iPSC)系,为获取组织的侵入性且有伦理争议的干预措施提供了一种替代方法。多能性iPSC实际上具有无限增殖能力,并能分化为身体的所有细胞类型。我们正在生成高质量、经过验证的NHP iPSC系,提供给科学界并促进他们的研究项目。我们使用非整合型附加体质粒载体系统从NHP皮肤活检样本中生成iPSC。在本章中,我们根据纳入记分卡的一组标记基因的既定标准和可测量限度,通过确认多能性及其分化为外胚层、中胚层和内胚层这三个胚层的倾向,来描述NHP iPSC系的验证。