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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
PMID34806042
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.
Sigma-Aldrich
Dimethyl sulfoxide, Hybri-Max™, sterile-filtered, BioReagent, suitable for hybridoma, ≥99.7%
Sigma-Aldrich
ESGRO® Recombinant Mouse LIF Protein, ESGRO Leukemia Inhibitory Factor (LIF) supplement for mouse ES cell culture. Each vial contains 10^7 units/ml.
Sigma-Aldrich
Fibronectin human, recombinant, expressed in HEK 293 cells, lyophilized powder, suitable for cell culture
Sigma-Aldrich
Bovine Serum Albumin, chromatographically purified, New Zealand origin, low endotoxin, suitable for cell culture, pH 7, ≥98%
Sigma-Aldrich
Dulbecco′s Phosphate Buffered Saline, Modified, without calcium chloride and magnesium chloride, liquid, sterile-filtered, suitable for cell culture