Establishment of Bovine-Induced Pluripotent Stem Cells

• Published in: International journal of molecular sciences Vol. 22; no. 19; p. 10489
• Main Authors: Su, Yue, Wang, Ling, Fan, Zhiqiang, Liu, Ying, Zhu, Jiaqi, Kaback, Deborah, Oudiz, Julia, Patrick, Tayler, Yee, Siu Pok, Tian, Xiuchun (Cindy), Polejaeva, Irina, Tang, Young
• Format: Journal Article
• Language: English
• Published: Basel MDPI AG 01.10.2021; MDPI
• Subjects: Antigens; bovine; Cattle; Cell proliferation; Cell self-renewal; Cloning; Colonies; Colonization; Cytology; differentiation; Efficiency; embryo aggregation; Embryo cells; Embryos; Fibroblasts; Genetic engineering; induced pluripotent stem cells (iPSCs); Karyotypes; Kinases; KLF4 protein; Lysine; Mesenchyme; Morphology; Nuclear transfer; Oct-4 protein; Pluripotency; reprogramming; Stem cell transplantation; Stem cells; Transcriptomes
• ISSN: 1422-0067; 1661-6596; 1422-0067
• DOI: 10.3390/ijms221910489

Pluripotent stem cells (PSCs) have been successfully developed in many species. However, the establishment of bovine-induced pluripotent stem cells (biPSCs) has been challenging. Here we report the generation of biPSCs from bovine mesenchymal stem cells (bMSCs) by overexpression of lysine-specific demethylase 4A (KDM4A) and the other reprogramming factors OCT4, SOX2, KLF4, cMYC, LIN28, and NANOG (KdOSKMLN). These biPSCs exhibited silenced transgene expression at passage 10, and had prolonged self-renewal capacity for over 70 passages. The biPSCs have flat, primed-like PSC colony morphology in combined media of knockout serum replacement (KSR) and mTeSR, but switched to dome-shaped, naïve-like PSC colony morphology in mTeSR medium and 2i/LIF with single cell colonization capacity. These cells have comparable proliferation rate to the reported primed- or naïve-state human PSCs, with three-germ layer differentiation capacity and normal karyotype. Transcriptome analysis revealed a high similarity of biPSCs to reported bovine embryonic stem cells (ESCs) and embryos. The naïve-like biPSCs can be incorporated into mouse embryos, with the extended capacity of integration into extra-embryonic tissues. Finally, at least 24.5% cloning efficiency could be obtained in nuclear transfer (NT) experiment using late passage biPSCs as nuclear donors. Our report represents a significant advance in the establishment of bovine PSCs.