Generation of induced pluripotent stem cells from Bornean orangutans

Chia-Jung Li; Chia-Chun Chang; Li-Kuang Tsai; Min Peng; Wei-Ni Lyu; Jane-Fang Yu; Mong-Hsun Tsai; Li-Ying Sung

doi:10.3389/fcell.2023.1331584

Generation of induced pluripotent stem cells from Bornean orangutans

Front Cell Dev Biol. 2024 Jan 5:11:1331584. doi: 10.3389/fcell.2023.1331584. eCollection 2023.

Authors

Chia-Jung Li^{1

2}, Chia-Chun Chang¹, Li-Kuang Tsai¹, Min Peng¹, Wei-Ni Lyu¹, Jane-Fang Yu³, Mong-Hsun Tsai¹, Li-Ying Sung^{1

4

5

6}

Affiliations

¹ Institute of Biotechnology, National Taiwan University, Taipei, Taiwan.
² Department of Obstetrics and Gynecology, Chang Gung Memorial Hospital, Linkou Branch, Chang Gung University, Taoyuan, Taiwan.
³ Conservation and Research Center, Taipei Zoo, Taipei, Taiwan.
⁴ Center for Developmental Biology and Regenerative Medicine, Taipei, Taiwan.
⁵ Center for Biotechnology, National Taiwan University, Taipei, Taiwan.
⁶ Agricultural Biotechnology Research Center, Academia Sinica, Taipei, Taiwan.

Abstract

Introduction: Orangutans, classified under the Pongo genus, are an endangered non-human primate (NHP) species. Derivation of induced pluripotent stem cells (iPSCs) represents a promising avenue for conserving the genetic resources of these animals. Earlier studies focused on deriving orangutan iPSCs (o-iPSCs) from Sumatran orangutans (Pongo abelii). To date, no reports specifically target the other Critically Endangered species in the Pongo genus, the Bornean orangutans (Pongo pygmaeus). Methods: Using Sendai virus-mediated Yamanaka factor-based reprogramming of peripheral blood mononuclear cells to generate iPSCs (bo-iPSCs) from a female captive Bornean orangutan. In this study, we evaluate the colony morphology, pluripotent markers, X chromosome activation status, and transcriptomic profile of the bo-iPSCs to demonstrate the pluripotency of iPSCs from Bornean orangutans. Results: The bo-iPSCs were successfully derived from Bornean orangutans, using Sendai virus-mediated Yamanaka factor-based reprogramming of peripheral blood mononuclear cells. When a modified 4i/L/A (m4i/L/A) culture system was applied to activate the WNT signaling pathway in these bo-iPSCs, the derived cells (m-bo-iPSCs) manifested characteristics akin to human naive pluripotent stem cells, including high expression levels of KLF17, DNMT3L, and DPPA3/5, as well as the X chromosome reactivation. Comparative RNA-seq analysis positioned the m-bo-iPSCs between human naive and formative pluripotent states. Furthermore, the m-bo-iPSCs express differentiation capacity into all three germlines, evidenced by controlled in vitro embryoid body formation assay. Discussion: Our work establishes a novel approach to preserve the genetic diversity of endangered Bornean orangutans while offering insights into primate stem cell pluripotency. In the future, derivation of the primordial germ cell-like cells (PGCLCs) from m-bo-iPSCs is needed to demonstrate the further specific application in species preservation and broaden the knowledge of primordial germ cell specification across species.

Keywords: Bornean orangutan; differentiation; endangered species; induced pluripotent stem cell; pluripotency; reprogramming.

Grants and funding

The author(s) declare financial support was received for the research, authorship, and/or publication of this article. This work was supported by grants from the Ministry of Science and Technology, Taiwan, R.O.C. (Grant Number MOST 109-2313-B-002-003-MY2 and MOST 111-2313-B-002-063) to L-YS and Chang Gung Memorial Hospital at Linkou, Taiwan, R.O.C., (CMRPG3L0171) to C-JL.