Characterization of the single-cell derived bovine induced pluripotent stem cells

Lixia Zhao; Zixin Wang; Jindun Zhang; Jian Yang; Xuefei Gao; Baojiang Wu; Gaoping Zhao; Siqin Bao; Shuxiang Hu; Pentao Liu; Xihe Li

doi:10.1016/j.tice.2017.05.005

Characterization of the single-cell derived bovine induced pluripotent stem cells

Tissue Cell. 2017 Oct;49(5):521-527. doi: 10.1016/j.tice.2017.05.005. Epub 2017 May 22.

Authors

Lixia Zhao¹, Zixin Wang², Jindun Zhang³, Jian Yang⁴, Xuefei Gao⁴, Baojiang Wu², Gaoping Zhao², Siqin Bao³, Shuxiang Hu¹, Pentao Liu⁴, Xihe Li⁵

Affiliations

¹ Research Center For Animal Genetic Resources of Mongolia Plateau, Inner Mongolia University, Hohhot, 010021, China; Inner Mongolia Saikexing Institute of Breeding and Reproductive Biotechnology in Domestic Animal, Hohhot, 011517, China.
² Inner Mongolia Saikexing Institute of Breeding and Reproductive Biotechnology in Domestic Animal, Hohhot, 011517, China.
³ Research Center For Animal Genetic Resources of Mongolia Plateau, Inner Mongolia University, Hohhot, 010021, China.
⁴ Wellcome Trust Sanger Institute, Wellcome Trust Genome Campus, Hinxton, Cambridgeshire CB10 1HH, UK.
⁵ Research Center For Animal Genetic Resources of Mongolia Plateau, Inner Mongolia University, Hohhot, 010021, China; Inner Mongolia Saikexing Institute of Breeding and Reproductive Biotechnology in Domestic Animal, Hohhot, 011517, China. Electronic address: lixh@life.imu.edu.cn.

PMID: 28720304
DOI: 10.1016/j.tice.2017.05.005

Abstract

Single-cell derived bovine induced pluripotent stem cells (iPSCs) were generated by the introduction of piggyBac transposons with CAG promoting transcription factors (Oct3/4, Sox2, Klf4 and cMyc). In the study, the bovine iPSCs colony from single cell could passage more than 50 passages after enzymatic dissociation into single cells. These bovine iPSCs cells kept the normal karyotype and displayed dome shaped clones similar to mouse embryonic stem cells. They showed pluripotency in many ways, including their expression of pluripotency markers, such as OCT3/4, NANOG, SOX2, SSEA1, SSEA4, and AP in immunofluorescence assay, Oct4, Nanog, Sox2, Klf4 and cMyc in RT-PCR. Additionally, single-cell derived bovine iPSCs formed embryoid bodies and teratomas that all subsequently gave rise to differentiated cells from all three embryonic germ layers. The results showed that our reprogramming method could obtain high efficiency single-cell cloning bovine iPSCs, and the efficiency of single cell cloning is 40%.

Keywords: Bovine iPSCs; Pluripotency; Single-cell; piggyBac.

MeSH terms

Animals
Cattle
Cell Culture Techniques
Cells, Cultured
Induced Pluripotent Stem Cells / cytology*
Kruppel-Like Factor 4