Abstract
BackgroundOrangutans 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.ResultsPrimary 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.ConclusionsWe 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.Electronic supplementary materialThe online version of this article (doi:10.1186/s13104-015-1567-0) contains supplementary material, which is available to authorized users.
Highlights
Orangutans are an endangered species whose natural habitats are restricted to the Southeast Asian islands of Borneo and Sumatra
IPSCs have been reported for small numbers of chimpanzees, bonobos, and gorillas using traditional Yamanaka reprogramming factors and retroviral vectors that integrated into the genome [18, 19]. induced pluripotent stem cell (iPSC) has been derived from two chimpanzees using reprogramming methods that do not involve the genomic integration of transgenes [20]
Derivation of candidate iPSCs from primary orangutan skin fibroblasts Primary skin fibroblast cultures previously derived from punch biopsies of the upper limbs of two Sumatran orangutans (KB10973, 29 year old male; KB10460, 43 year old female) [33] were obtained from the Zoological Society of San Diego
Summary
IPSCs can be expanded indefinitely and differentiated into cells from all three germ layers This is highlighted by ongoing efforts to develop methods to differentiate iPSCs from endangered species into germ cells for the in vitro production of embryos that can be implanted into surrogate females of a related existing species [12]. This would further enhance the value of cryopreserved cells banks, such as the Frozen Zoo© at the Zoological Society of San Diego [13, 14]. Despite these reports involving African great apes as well as non-endangered primates used in biomedical research [21], the derivation of iPSCs from orangutans has not been reported to date
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