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Induction of human trophoblast stem cells

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Abstract

Cell reprogramming has allowed unprecedented access to human development, from virtually any genome. However, reprogramming yields pluripotent stem cells that can differentiate into all cells that form a fetus, but not extraembryonic annexes. Therefore, a cellular model allowing study of placental development from a broad genomic repertoire is lacking. Here, we describe an optimized protocol to reprogram somatic cells into human induced trophoblast stem cells (hiTSCs) and convert pluripotent stem cells into human converted TSCs (hcTSCs). This protocol enables much-needed genome-specific placental disease modeling. We also detail extravillous trophoblast and syncytiotrophoblast differentiation protocols from hiTSCs and hcTSCs, a necessary step to validate these cells. In total, this protocol takes 4 months and requires advanced cell culture skills, comparable to those necessary for somatic cell reprogramming into human induced pluripotent stem cells.

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Fig. 1: Morphological changes associated with hiTSC reprogramming and conversion.
Fig. 2: Controlling outcomes of reprogramming and conversion assays.
Fig. 3: Molecular validation of induced and converted hTSCs.
Fig. 4: Functional validation of hi/cTSC differentiation potential.

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Data availability

All datasets used in this paper were uploaded to the European Nucleotide Archive (https://www.ebi.ac.uk/ena/browser/view/PRJEB34037?show=reads), as specified in our previous paper5.

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Acknowledgements

G.C. is a recipient of an ‘INSERM-Région Pays de la Loire’ fellowship. We acknowledge the MicroPICell, GenoA, BIRD and iPSC core facilities, all supported by Biogenouest and IBiSA, for the use of their resources and technical support. MicroPICell is a member of the national infrastructure France-Bioimaging (ANR-10-INBS-04). BIRD is a member of Institut Français de Bioinformatique (IFB) (ANR-11-INBS-0013). We thank M. Narimatsu for the tips regarding coverslip preparation.

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G.C. and L.D. designed experiments, and G.C. conducted them. G.C. and L.D. wrote the manuscript.

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Correspondence to Laurent David.

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Nature Protocols thanks Berthold Huppertz and Michael Roberts for their contribution to the peer review of this work.

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Key reference using this protocol

Castel, G. et al. Cell Rep. 33, 108419 (2020): https://doi.org/10.1016/j.celrep.2020.108419

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Castel, G., David, L. Induction of human trophoblast stem cells. Nat Protoc 17, 2760–2783 (2022). https://doi.org/10.1038/s41596-022-00744-0

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