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Merck

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
RESUMO

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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Dimetilsulfóxido, Hybri-Max, sterile-filtered, BioReagent, suitable for hybridoma, ≥99.7%
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Solução salina tamponada com fosfato, 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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Proteína LIF recombinante de camundongo ESGRO®, ESGRO Leukemia Inhibitory Factor (LIF) supplement for mouse ES cell culture. Each vial contains 10^7 units/ml.
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Paraformaldehyde, powder, 95%
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Piruvato de sódio, powder, BioXtra, suitable for mouse embryo cell culture
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Albumina sérica 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