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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).

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Sigma-Aldrich
Dimetil solfossido, Hybri-Max, sterile-filtered, BioReagent, suitable for hybridoma, ≥99.7%
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Tampone fosfato salino di Dulbecco, 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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Proteina LIF ricombinante di topo 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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Sodium pyruvate, powder, BioXtra, suitable for mouse embryo cell culture
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Fibronectina, recombinant, expressed in HEK 293 cells, lyophilized powder, suitable for cell culture