通过piggyBac转位重编程因子生成狒狒诱导多能干细胞。

Q2 Agricultural and Biological Sciences Primate Biology Pub Date : 2019-07-29 eCollection Date: 2019-01-01 DOI:10.5194/pb-6-75-2019
Ignacio Rodriguez-Polo, Michael Stauske, Alexander Becker, Iris Bartels, Ralf Dressel, Rüdiger Behr
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引用次数: 0

摘要

利用胚胎干细胞或诱导多能干细胞进行再生治疗的临床应用指日可待。近年来取得的进展鼓励研究人员解决尚未解决的问题,以便最终将实验性细胞替代疗法应用于患者。为实现这一目标,需要对转化相关的动物模型进行研究,以迈出临床的最后一步。在这种情况下,狒狒(Papio anubis)可能是测试细胞替代疗法的一种有价值的非人灵长类动物(NHP)模型,因为它与人类的进化关系密切,而且体型庞大。在这项研究中,我们描述了利用piggyBac转座子系统对成年狒狒皮肤成纤维细胞进行重编程的过程。通过转座子介导的六种重编程因子的过表达,我们产生了五个狒狒诱导多能干细胞(iPSC)系。我们从碱性磷酸酶活性、多能因子表达分析、畸胎瘤形成潜能和核型等方面对这些 iPSC 株进行了鉴定。此外,在与小鼠胚胎成纤维细胞进行初始共培养后,我们能够使 iPSC 株系适应无饲养者条件。总之,我们建立了从成年狒狒成纤维细胞生成 iPSC 的稳健而高效的方案。
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Baboon induced pluripotent stem cell generation by piggyBac transposition of reprogramming factors.

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 (Papio anubis) 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 piggyBac 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.

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来源期刊
Primate Biology
Primate Biology Agricultural and Biological Sciences-Animal Science and Zoology
CiteScore
1.90
自引率
0.00%
发文量
8
审稿时长
13 weeks
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