Rodriguez-Polo Ignacio, Stauske Michael, Becker Alexander, Bartels Iris, Dressel Ralf, Behr Rüdiger
Research Platform Degenerative Diseases, German Primate Center - Leibniz Institute for Primate Research, Kellnerweg 4, 37077 Göttingen, Germany.
German Center for Cardiovascular Research (DZHK), Partner site, Göttingen, Germany.
Primate Biol. 2019 Jul 29;6(2):75-86. doi: 10.5194/pb-6-75-2019. eCollection 2019.
Clinical application of regenerative therapies using embryonic or induced pluripotent stem cells is within reach. Progress made during recent years has encouraged researchers to address remaining open questions in order to finally translate experimental cell replacement therapies into application in patients. To achieve this, studies in translationally relevant animal models are required to make the final step to the clinic. In this context, the baboon () may represent a valuable nonhuman primate (NHP) model to test cell replacement therapies because of its close evolutionary relationship to humans and its large body size. In this study, we describe the reprogramming of adult baboon skin fibroblasts using the transposon system. Via transposon-mediated overexpression of six reprogramming factors, we generated five baboon induced pluripotent stem cell (iPSC) lines. The iPSC lines were characterized with respect to alkaline phosphatase activity, pluripotency factor expression analysis, teratoma formation potential, and karyotype. Furthermore, after initial cocultivation with mouse embryonic fibroblasts, we were able to adapt iPSC lines to feeder-free conditions. In conclusion, we established a robust and efficient protocol for iPSC generation from adult baboon fibroblasts.
使用胚胎干细胞或诱导多能干细胞的再生疗法的临床应用已指日可待。近年来取得的进展鼓励研究人员解决悬而未决的问题，以便最终将实验性细胞替代疗法转化应用于患者。要实现这一目标，需要在与转化相关的动物模型中进行研究，以迈出迈向临床应用的最后一步。在这种情况下，狒狒（ ）可能是一种有价值的非人灵长类动物（NHP）模型，可用于测试细胞替代疗法，因为它与人类有着密切的进化关系且体型较大。在本研究中，我们描述了使用转座子系统对成年狒狒皮肤成纤维细胞进行重编程。通过转座子介导的六种重编程因子的过表达，我们生成了五条狒狒诱导多能干细胞（iPSC）系。对iPSC系进行了碱性磷酸酶活性、多能性因子表达分析、畸胎瘤形成潜力和核型等方面的特征鉴定。此外，在与小鼠胚胎成纤维细胞初步共培养后，我们能够使iPSC系适应无饲养层培养条件。总之，我们建立了一种从成年狒狒成纤维细胞生成iPSC的稳健且高效的方案。
