Derivation of Haploid Trophoblast Stem Cells via Conversion In Vitro

Keli Peng; Xu Li; Congyu Wu; Yuna Wang; Jian Yu; Jinxin Zhang; Qian Gao; Wenhao Zhang; Qian Zhang; Yong Fan; Yang Yu; Ling Shuai

doi:10.1016/j.isci.2018.12.014

Derivation of Haploid Trophoblast Stem Cells via Conversion In Vitro

iScience. 2019 Jan 25:11:508-518. doi: 10.1016/j.isci.2018.12.014. Epub 2019 Jan 8.

Authors

Keli Peng¹, Xu Li¹, Congyu Wu², Yuna Wang¹, Jian Yu³, Jinxin Zhang¹, Qian Gao⁴, Wenhao Zhang¹, Qian Zhang¹, Yong Fan⁵, Yang Yu⁶, Ling Shuai⁷

Affiliations

¹ State Key Laboratory of Medicinal Chemical Biology and College of Pharmacy, Nankai University, Tianjin 300350, China.
² Department of Neurological Surgery, Feinberg School of Medicine, Northwestern University, Chicago 60611, USA.
³ Swiss Federal Institute of Technology Zurich, Department of Biology, Institute of Molecular Health Sciences, Chair of RNAi and Genome Integrity, Zurich 8093, Switzerland.
⁴ State Key Laboratory of Medicinal Chemical Biology and College of Pharmacy, Nankai University, Tianjin 300350, China; Department of Gynecology and Obstetrics, Peking University Third Hospital, Beijing 100191, China.
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.
⁶ Department of Gynecology and Obstetrics, Peking University Third Hospital, Beijing 100191, China.
⁷ State Key Laboratory of Medicinal Chemical Biology and College of Pharmacy, Nankai University, Tianjin 300350, China. Electronic address: lshuai@nankai.edu.cn.

Abstract

Owing to their single genome, haploid cells are powerful to uncover unknown genes by performing genetic screening in mammals. However, no haploid cell line from an extraembryonic lineage has been achieved yet, which limits the application of haploid cells in placental genetic screening. Here, we show that overexpression of Cdx2 can convert haploid embryonic stem cells to trophoblast stem cells (TSCs). p53 deletion reduces diploidization during the conversion and guarantees the generation of haploid-induced TSCs (haiTSCs). haiTSCs not only share the same molecular characterization with trophoderm-derived TSCs but also possess multipotency to placental lineages in various procedures. In addition, haiTSCs can maintain haploidy in the long term, assisted by periodic sorting and with reliance on FGF4 and heparin. Finally, we perform piggyBac-mediated high-throughput mutation in haiTSCs and use them in trophoblast lineage genetic screening. Deep sequencing analysis and validation experiments prove that Htra1 is a blocker for spongiotrophoblast specification.

Keywords: Cell Biology; Genomics; Stem Cells Research.