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Rapid generation of murine haploid-induced trophoblast stem cells via a Tet-on system.

STAR protocols (2021-11-23)
Mei Xu, Wenhao Zhang, Mengyang Geng, Yiding Zhao, Shengyi Sun, Qian Gao, Yan Liu, Ling Shuai
ABSTRACT

Haploid trophoblast stem cells (TSCs) are advanced in studying placental development for their placental precursor and homozygous features. Here, we describe how to generate haploid-induced TSCs (haiTSCs) from haploid embryonic stem cells with a Tet-on system. Our haiTSCs can maintain haploidy long-term and can produce genome-wide mutants combined with transposons. It is promising in high-throughput genetic screening of trophoblast-specific modulators. For complete details on the use and execution of this protocol, please refer to Peng et al. (2019).

MATERIALS
Product Number
Brand
Product Description

Millipore
Hygromycin B, Streptomyces sp., Cell Culture-Tested - CAS 31282-04-9 - Calbiochem, Unique aminoglycoside antibiotic that inhibits the growth of prokaryotic (bacteria) and eukaryotic microorganisms (yeasts) and mammalian cells.
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Triton X-100, for molecular biology
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Dimethyl sulfoxide, Hybri-Max, sterile-filtered, BioReagent, suitable for hybridoma, ≥99.7%
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Dulbecco′s Phosphate Buffered Saline, Modified, without calcium chloride and magnesium chloride, liquid, sterile-filtered, suitable for cell culture