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Merck
  • 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
摘要

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).

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Sigma-Aldrich
二甲基亚砜, Hybri-Max, sterile-filtered, BioReagent, suitable for hybridoma, ≥99.7%
Sigma-Aldrich
杜氏磷酸盐缓冲盐水, Modified, without calcium chloride and magnesium chloride, liquid, sterile-filtered, suitable for cell culture
Sigma-Aldrich
Triton X-100, for molecular biology
Sigma-Aldrich
ESGRO®重组小鼠LIF蛋白, ESGRO Leukemia Inhibitory Factor (LIF) supplement for mouse ES cell culture. Each vial contains 10^7 units/ml.
Sigma-Aldrich
多聚甲醛, powder, 95%
Sigma-Aldrich
丙酮酸钠, powder, BioXtra, suitable for mouse embryo cell culture
Sigma-Aldrich
牛血清白蛋白 来源于牛血清, chromatographically purified, New Zealand origin, low endotoxin, suitable for cell culture, pH 7, ≥98%
Sigma-Aldrich
人纤维连接蛋白, recombinant, expressed in HEK 293 cells, lyophilized powder, suitable for cell culture
Millipore
潮霉素B,来源于链霉菌属,经细胞培养测试, Unique aminoglycoside antibiotic that inhibits the growth of prokaryotic (bacteria) and eukaryotic microorganisms (yeasts) and mammalian cells.