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

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

Sigma-Aldrich
Paraformaldehyde, powder, 95%
Sigma-Aldrich
Sodium pyruvate, powder, BioXtra, suitable for mouse embryo cell culture
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Bovine Serum Albumin, chromatographically purified, New Zealand origin, low endotoxin, suitable for cell culture, pH 7, ≥98%
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Dulbecco′s Phosphate Buffered Saline, Modified, without calcium chloride and magnesium chloride, liquid, sterile-filtered, suitable for cell culture
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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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ESGRO® Recombinant Mouse LIF Protein, ESGRO Leukemia Inhibitory Factor (LIF) supplement for mouse ES cell culture. Each vial contains 10^7 units/ml.
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Fibronectin human, recombinant, expressed in HEK 293 cells, lyophilized powder, suitable for cell culture