BCL2 is a major regulator of haploidy maintenance in murine embryonic stem cells

• Published in: Cell proliferation Vol. 56; no. 12; pp. e13498 - n/a
• Main Authors: Sun, Shengyi, Zhao, Qin, Zhao, Yiding, Geng, Mengyang, Wang, Qing, Gao, Qian, Zhang, Xiao‐Ou, Zhang, Wenhao, Shuai, Ling
• Format: Journal Article
• Language: English
• Published: England John Wiley & Sons, Inc 01.12.2023; John Wiley and Sons Inc
• Subjects: Antibodies; Bcl-2 protein; Cell culture; Cell cycle; Cell death; Cell differentiation; Cell division; Cell lines; Differentiation; Drug development; Embryo cells; Embryogenesis; Fetuses; Genes; Genetic screening; Haploidy; Laboratory animals; Maintenance; Medical research; Membranes; Original; Plasmids; Polymerase chain reaction; Stem cells; Teratoma; Transcriptomes
• ISSN: 0960-7722; 1365-2184
• DOI: 10.1111/cpr.13498

Mammalian haploid cells are important resources for forward genetic screening and are important in genetic medicine and drug development. However, the self‐diploidization of murine haploid embryonic stem cells (haESCs) during daily culture or differentiation jeopardizes their use in genetic approaches. Here, we show that overexpression (OE) of an antiapoptosis gene, BCL2, in haESCs robustly ensures their haploidy maintenance in various situations, even under strict differentiation in vivo (embryonic 10.5 chimeric fetus or 21‐day teratoma). Haploid cell lines of many lineages, including epiblasts, trophectodermal lineages, and neuroectodermal lineages, can be easily derived by the differentiation of BCL2‐OE haESCs in vitro. Transcriptome analysis revealed that BCL2‐OE activates another regulatory gene, Has2, which is also sufficient for haploidy maintenance. Together, our findings provide an effective and secure strategy to reduce diploidization during differentiation, which will contribute to the generation of haploid cell lines of the desired lineage and related genetic screening. BCL2‐OE haESCs present reduced diploidization and sustain haploidy stably in differentiated cells, with an intact genome. A BCL2‐related gene, Has2, is also sufficient to haploidy maintenance. These findings provide a premium and secure strategy for haploidy maintenance of mouse haESCs, guaranteeing their advantages in genetic approaches.