Abstract
Currently, human trophoblast stem cell (hTSC) is considered to be the most promising laboratory model stimulating trophoblast criteria. Our group has established hTSCs allowing differentiation to syncytiotrophoblast (STB) and extravillous trophoblast (EVT). Further, hTSC-based three-dimensional (3D) trophoblast organoid (hTSC-organoid) provides a transformative model for studying human placental development and the interaction between trophoblast and maternal environment. Here, we present a protocol to obtain different types of placental trophoblast cells and trophoblast organoids using hTSCs. The generation of hTSC-organoids takes 6 days. hTSC-organoids permit passaging and can differentiate into EVT lineage.
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References
Maltepe E, Fisher SJ (2015) Placenta: the forgotten organ. Annu Rev Cell Dev Biol 31:523–552
Delorme-Axford E, Sadovsky Y, Coyne CB (2014) The placenta as a barrier to viral infections. Annu Rev Virol 1(1):133–146
Bonnin A, Goeden N, Chen K, Wilson ML, King J, Shih JC, Blakely RD, Deneris ES, Levitt P (2011) A transient placental source of serotonin for the fetal forebrain. Nature 472(7343):347–350
Eriksson JG, Kajantie E, Thornburg KL, Osmond C, Barker DJP (2011) Mother’s body size and placental size predict coronary heart disease in men. Eur Heart J 32(18):2297–2303
Benirschke’s pathology of the human placenta (2022)
Shahbazi MN, Zernicka-Goetz M (2018) Deconstructing and reconstructing the mouse and human early embryo. Nat Cell Biol 20(8):878–887
Shahbazi MN (2020) Mechanisms of human embryo development: from cell fate to tissue shape and back. Development 147(14):dev190629
Okae H, Toh H, Sato T, Hiura H, Takahashi S, Shirane K, Kabayama Y, Suyama M, Sasaki H, Arima T (2018) Derivation of human trophoblast stem cells. Cell stem cell 22(1):50–63. e56
Wang Y, Wu H, Jiang X, Jia L, Wang M, Rong Y, Chen S, Wang Y, Xiao Z, Liang X et al (2022) LMNA determines nuclear morphology during syncytialization of human trophoblast stem cells. Front Cell Dev Biol 10:836390
Xiao Z, Yan L, Liang X, Wang H (2020) Progress in deciphering trophoblast cell differentiation during human placentation. Curr Opin Cell Biol 67:86–91
Liu Y, Fan X, Wang R, Lu X, Dang YL, Wang H, Lin HY, Zhu C, Ge H, Cross JC et al (2018) Single-cell RNA-seq reveals the diversity of trophoblast subtypes and patterns of differentiation in the human placenta. Cell Res 28(8):819–832
Lu X, Wang R, Zhu C, Wang H, Lin HY, Gu Y, Cross JC, Wang H (2018) Fine-tuned and cell-cycle-restricted expression of fusogenic protein syncytin-2 maintains functional placental syncytia. Cell Rep 23(13):3979
Chang WL, Liu YW, Dang YL, Jiang XX, Xu H, Huang X, Wang YL, Wang H, Zhu C, Xue LQ et al (2018) PLAC8, a new marker for human interstitial extravillous trophoblast cells, promotes their invasion and migration. Development 145(2):dev148932
Malassiné A, Cronier L (2002) Hormones and human trophoblast differentiation: a review. Endocrine 19(1):3–11
Genbacev O, Donne M, Kapidzic M, Gormley M, Lamb J, Gilmore J, Larocque N, Goldfien G, Zdravkovic T, McMaster MT et al (2011) Establishment of human trophoblast progenitor cell lines from the chorion. Stem Cells 29(9):1427–1436
Zdravkovic T, Nazor KL, Larocque N, Gormley M, Donne M, Hunkapillar N, Giritharan G, Bernstein HS, Wei G, Hebrok M et al (2015) Human stem cells from single blastomeres reveal pathways of embryonic or trophoblast fate specification. Development 142(23):4010–4025
Xu RH, Chen X, Li DS, Li R, Addicks GC, Glennon C, Zwaka TP, Thomson JA (2002) BMP4 initiates human embryonic stem cell differentiation to trophoblast. Nat Biotechnol 20(12):1261–1264
Wei Y, Wang T, Ma L, Zhang Y, Zhao Y, Lye K, Xiao L, Chen C, Wang Z, Ma Y et al (2021) Efficient derivation of human trophoblast stem cells from primed pluripotent stem cells. Sci Adv 7(33):eabf4416
Io S, Kabata M, Iemura Y, Semi K, Morone N, Minagawa A, Wang B, Okamoto I, Nakamura T, Kojima Y et al (2021) Capturing human trophoblast development with naive pluripotent stem cells in vitro. Cell Stem Cell 28(6):1023–1039.e1013
Pattillo RA, Gey GO (1968) The establishment of a cell line of human hormone-synthesizing trophoblastic cells in vitro. Cancer Res 28(7):1231–1236
Kohler PO, Bridson WE (1971) Isolation of hormone-producing clonal lines of human choriocarcinoma. J Clin Endocrinol Metab 32(5):683–687
Barnea ER, Ashkenazy R, Tal Y, Kol S, Sarne Y (1991) Effect of beta-endorphin on human chorionic gonadotrophin secretion by placental explants. Hum Reprod 6(9):1327–1331
Turco MY, Gardner L, Kay RG, Hamilton RS, Prater M, Hollinshead MS, McWhinnie A, Esposito L, Fernando R, Skelton H et al (2018) Trophoblast organoids as a model for maternal-fetal interactions during human placentation. Nature 564(7735):263–267
Haider S, Meinhardt G, Saleh L, Kunihs V, Gamperl M, Kaindl U, Ellinger A, Burkard TR, Fiala C, Pollheimer J et al (2018) Self-renewing trophoblast organoids recapitulate the developmental program of the early human placenta. Stem Cell Rep 11(2):537–551
Acknowledgments
This work was supported by National Natural Science Foundation of China (82192870) and National Key R&D Program of China (2021YFC2700302, 2018YFE0201103).
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Wu, H., Wang, Y., Wang, H. (2022). Generation of Human Trophoblast Stem Cell-Dependent Placental In Vitro Models. In: Zernicka-Goetz, M., Turksen, K. (eds) Embryo Models In Vitro. Methods in Molecular Biology, vol 2767. Humana, New York, NY. https://doi.org/10.1007/7651_2022_463
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DOI: https://doi.org/10.1007/7651_2022_463
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