Derivation of induced pluripotent stem cells from orangutan skin fibroblasts

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.

Functional Divergence of the Nuclear ReceptorNR2C1as a Modulator of Pluripotentiality During Hominid Evolution

“…Finally, analysis of NR2C1 expression in the mouse embryo illustrates that it is robustly expressed in placodally derived neuroectodermal stem cells and presumptive neural-crest-derived mesenchymal stem cells . Further investigation as to whether this gene can regulate self-renewal and/or pluripotentiality in populations of stem cells other than ES cells such as in cultures of neural stem cells, or in iPSCs generated from nonhuman primates (Marchetto et al 2013;Wunderlich et al 2014;Gallego Romero et al 2015;Ramaswamy et al 2015) is warranted. iPSCs are an incredibly useful tool and provide a great deal of insight into questions of pluripotentiality and differentiation (Takahashi and Yamanaka 2006); however, some questions remain regarding whether epigenetic reprogramming during the iPSC protocol results in full pluripotency potential (Bilic and Belmonte 2012;Robinton and Daley 2012;Halevy and Urbach 2014).…”

Gametes from stem cells: Status and applications in animal reproduction

“…On the other hand, iPSCs are equivalent to ESCs in multiple aspects, including morphology, growth, gene expression, teratoma formation and contribution to germline chimeras. Lines of iPSCs have been reported for a number of livestock species, including cattle, pig, sheep, horse, goat, dog and some endangered species including drill monkeys, white rhinoceros, orangutan and snow leopard (Baird, Barsby, & Guest, ; Breton et al, ; Chu et al, ; Friedrich Ben‐Nun et al, ; Fujishiro et al, ; Han et al, ; Hildebrandt et al, ; Nagy et al, ; Ramaswamy et al, ; Sartori et al, ). In general, reprogramming factors were delivered into embryonic or foetal fibroblasts by viral vectors, and pluripotency was determined by formation of teratomas.…”

Applying iPSCs for Preserving Endangered Species and Elucidating the Evolution of Mammalian Sex Determination

“…Recently, generation of iPSCs from endangered species was achieved with the orangutan, drill, and northern white rhinoceros . Because all exhibited prime pluripotency, it seems difficult that they contributed to host embryos as a chimera.…”