Derivation of induced pluripotent stem cells from orangutan skin fibroblasts.
Derivation of induced pluripotent stem cells from orangutan skin fibroblasts.
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DOI：
10.1186/s13104-015-1567-0
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发表时间：
2015-10-16
影响因子：
1.8
通讯作者：
Hacia JG
中科院分区：
文献类型：
--
作者：
Ramaswamy K;Yik WY;Wang XM;Oliphant EN;Lu W;Shibata D;Ryder OA;Hacia JG
猩猩是一种濒危物种，其自然栖息地仅限于东南亚的婆罗洲和苏门答腊岛。沿着非洲大猿，猩猩是人类现存的近亲之一。为了潜在的物种保护和功能基因组学研究，我们从圈养猩猩的冻存体细胞中获得诱导多能干细胞（iPSC）。用表达人OCT 4、SOX 2、KLF 4和c-MYC因子的逆转录病毒载体转导来自两只苏门答腊猩猩的原代皮肤成纤维细胞。通过全局基因表达和DNA拷贝数分析来表征候选猩猩iPSC。所有这些都与多能性一致，并且没有提供大的基因组插入或缺失的证据。此外，猩猩iPSC能够通过胚状体分化试验在体外产生来自所有三个胚层的细胞，并通过免疫受损小鼠中的畸胎瘤形成在体内产生细胞。我们证明了猩猩皮肤成纤维细胞能够被重编程为具有标志性分子特征和分化潜力的iPSC。我们认为，在基因组资源库中重新编程猩猩体细胞可以为推进与物种保护工作相关的辅助生殖技术提供新的机会。此外，猩猩iPSC可以用于研究人类、非洲巨猿和/或猩猩谱系中发生的基因组变化的表型相关性。这为猩猩细胞培养模型提供了机会，否则由于获得原代细胞所需程序的侵入性，这些模型不可能从活体供体中开发出来。本文的在线版本（doi：10.1186/s13104-015-1567-0）包含补充材料，可供授权用户使用。
Orangutans are an endangered species whose natural habitats are restricted to the Southeast Asian islands of Borneo and Sumatra. Along with the African great apes, orangutans are among the closest living relatives to humans. For potential species conservation and functional genomics studies, we derived induced pluripotent stem cells (iPSCs) from cryopreserved somatic cells obtained from captive orangutans. Primary skin fibroblasts from two Sumatran orangutans were transduced with retroviral vectors expressing the human OCT4, SOX2, KLF4, and c-MYC factors. Candidate orangutan iPSCs were characterized by global gene expression and DNA copy number analysis. All were consistent with pluripotency and provided no evidence of large genomic insertions or deletions. In addition, orangutan iPSCs were capable of producing cells derived from all three germ layers in vitro through embryoid body differentiation assays and in vivo through teratoma formation in immune-compromised mice. We demonstrate that orangutan skin fibroblasts are capable of being reprogrammed into iPSCs with hallmark molecular signatures and differentiation potential. We suggest that reprogramming orangutan somatic cells in genome resource banks could provide new opportunities for advancing assisted reproductive technologies relevant for species conservation efforts. Furthermore, orangutan iPSCs could have applications for investigating the phenotypic relevance of genomic changes that occurred in the human, African great ape, and/or orangutan lineages. This provides opportunities for orangutan cell culture models that would otherwise be impossible to develop from living donors due to the invasive nature of the procedures required for obtaining primary cells. The online version of this article (doi:10.1186/s13104-015-1567-0) contains supplementary material, which is available to authorized users.