- 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
PMID34806042
RESUMEN
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).
MATERIALES
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Sigma-Aldrich
Dimetilsulfóxido, Hybri-Max™, sterile-filtered, BioReagent, suitable for hybridoma, ≥99.7%
Sigma-Aldrich
Disolución salina tamponada con fosfatos de Dulbecco, Modified, without calcium chloride and magnesium chloride, liquid, sterile-filtered, suitable for cell culture
Sigma-Aldrich
Proteína LIF recombinante de ratón ESGRO®, ESGRO Leukemia Inhibitory Factor (LIF) supplement for mouse ES cell culture. Each vial contains 10^7 units/ml.
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Seroalbúmina bovina, chromatographically purified, New Zealand origin, low endotoxin, suitable for cell culture, pH 7, ≥98%
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Fibronectina, recombinant, expressed in HEK 293 cells, lyophilized powder, suitable for cell culture
Millipore
Hygromycin B, Streptomyces sp., Cell Culture-Tested, Unique aminoglycoside antibiotic that inhibits the growth of prokaryotic (bacteria) and eukaryotic microorganisms (yeasts) and mammalian cells.