Inhibition of Apoptosis Reduces Diploidization of Haploid Mouse Embryonic Stem Cells during Differentiation

Wenhao Zhang¹, Yaru Tian¹, Qian Gao², Xu Li¹, Yanni Li³, Jinxin Zhang¹, Chunmeng Yao¹, Yuna Wang¹, Haoyu Wang¹, Yiding Zhao¹, Qian Zhang¹, Luyuan Li¹, Yang Yu⁴, Yong Fan⁵, Ling Shuai⁶

Affiliations

¹ State Key Laboratory of Medicinal Chemical Biology and College of Pharmacy, Nankai University, Tianjin 300350, China.
² State Key Laboratory of Medicinal Chemical Biology and College of Pharmacy, Nankai University, Tianjin 300350, China; Reproductive Medical Center, Department of Gynecology and Obstetrics, Clinical Stem Cell Research Center, Peking University Third Hospital, Beijing 100191, China.
³ Key Laboratory for Major Obstetric Diseases of Guangdong Province, Key Laboratory of Reproduction and Genetics of Guangdong Higher Education Institutes, The Third Affiliated Hospital of Guangzhou Medical University, Guangzhou 510150, China.
⁴ Reproductive Medical Center, Department of Gynecology and Obstetrics, Clinical Stem Cell Research Center, Peking University Third Hospital, Beijing 100191, China. Electronic address: yuyang5012@hotmail.com.
⁵ Key Laboratory for Major Obstetric Diseases of Guangdong Province, Key Laboratory of Reproduction and Genetics of Guangdong Higher Education Institutes, The Third Affiliated Hospital of Guangzhou Medical University, Guangzhou 510150, China. Electronic address: yongfan011@gzhmu.edu.cn.
⁶ State Key Laboratory of Medicinal Chemical Biology and College of Pharmacy, Nankai University, Tianjin 300350, China. Electronic address: lshuai@nankai.edu.cn.

PMID: 32502463
PMCID: PMC7363743
DOI: 10.1016/j.stemcr.2020.05.004

Free PMC article

Abstract

Phenotypes of haploid embryonic stem cells (haESCs) are dominant for recessive traits in mice. However, one major obstacle to their use is self-diploidization in daily culture. Although haESCs maintain haploidy well by deleting p53, whether they can sustain haploidy in differentiated status and the mechanism behind it remain unknown. To address this, we induced p53-deficient haESCs into multiple differentiated lineages maintain haploid status in vitro. Haploid cells also remained in chimeric embryos and teratomas arising from p53-null haESCs. Transcriptome analysis revealed that apoptosis genes were downregulated in p53-null haESCs compared with that in wild-type haESCs. Finally, we knocked out p73, another apoptosis-related gene, and observed stabilization of haploidy in haESCs. These results indicated that the main mechanism of diploidization was apoptosis-related gene-triggered cell death in haploid cell cultures. Thus, we can derive haploid somatic cells by manipulating the apoptosis gene, facilitating genetic screens of lineage-specific development.

Keywords: apoptosis; haESCs; haploidy; p53; p73; self-diploidization.

Stem Cell Reports. 2020 Jul 14;15(1):185-197.doi: 10.1016/j.stemcr.2020.05.004. Epub 2020 Jun 4. (IF:7.294).

Inhibition of Apoptosis Reduces Diploidization of Haploid Mouse Embryonic Stem Cells during Differentiation

Abstract

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