{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,12]],"date-time":"2026-09-12T14:44:19Z","timestamp":1789224259460,"version":"build-2803163510"},"reference-count":51,"publisher":"Wiley","issue":"10","license":[{"start":{"date-parts":[[2023,10,3]],"date-time":"2023-10-03T00:00:00Z","timestamp":1696291200000},"content-version":"vor","delay-in-days":2,"URL":"http:\/\/creativecommons.org\/licenses\/by-nc-nd\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Current Protocols"],"published-print":{"date-parts":[[2023,10]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:p>We previously established a trophoblast differentiation protocol from primed human pluripotent stem cells (PSC). To induce this lineage, we use a combination of Bone Morphogenetic Protein\u20104 (BMP4) and the WNT inhibitor IWP2. This protocol has enabled us to obtain a pure population of trophectoderm (TE)\u2010like cells that could subsequently be terminally differentiated into syncytiotrophoblasts (STB) and extravillous trophoblasts (EVT). However, the resulting TE\u2010like cells could only be terminally differentiated to a variable mixture of STB and EVT, with a bias toward the STB lineage. Recently, methods have been developed for derivation and culture of self\u2010renewing human trophoblast stem cells (TSC) from human embryos and early gestation placental tissues. These primary TSCs were further able to differentiate into either STB or EVT with high efficiency using the lineage specific differentiation protocols. Based partly on these protocols, we have developed methods for establishing self\u2010renewing TSC\u2010like cells from PSC, and for efficient lineage\u2010specific terminal differentiation. Here, we describe in detail the protocols to derive and maintain PSC\u2010TSC, from both embryonic stem cells (ESC) and patient\u2010derived induced pluripotent stem cells (iPSC), and their subsequent terminal differentiation to STB and EVT. \u00a9 2023 The Authors. Current Protocols published by Wiley Periodicals LLC.<\/jats:p><jats:p>This article was corrected on 14 April 2024. See the end of the full text for details.<\/jats:p><jats:p><jats:bold>Basic Protocol 1<\/jats:bold>: Trophoblast Differentiation into TE\u2010like Cells<\/jats:p><jats:p><jats:bold>Basic Protocol 2<\/jats:bold>: Conversion of PSC\u2010Derived TE\u2010like Cells to TSC<\/jats:p><jats:p><jats:bold>Basic Protocol 3<\/jats:bold>: Passaging PSC\u2010Derived TSC in iCTB Complete Medium<\/jats:p><jats:p><jats:bold>Basic Protocol 4<\/jats:bold>: STB Differentiation from PSC\u2010derived TSC<\/jats:p><jats:p><jats:bold>Basic Protocol 5<\/jats:bold>: EVT Differentiation from PSC\u2010derived TSC<\/jats:p><jats:p><jats:bold>Support Protocol 1<\/jats:bold>: Geltrex\u2010coated tissue culture plate preparation<\/jats:p><jats:p><jats:bold>Support Protocol 2<\/jats:bold>: Collagen IV\u2010coated tissue culture plate preparation<\/jats:p><jats:p><jats:bold>Support Protocol 3<\/jats:bold>: Fibronectin\u2010coated tissue culture plate preparation<\/jats:p>","DOI":"10.1002\/cpz1.875","type":"journal-article","created":{"date-parts":[[2023,10,4]],"date-time":"2023-10-04T08:38:21Z","timestamp":1696408701000},"source":"Crossref","is-referenced-by-count":8,"title":["Current Strategies of Modeling Human Trophoblast Using Human Pluripotent Stem Cells <i>in vitro<\/i>"],"prefix":"10.1002","volume":"3","author":[{"given":"Virginia Chu","family":"Cheung","sequence":"first","affiliation":[{"name":"Department of Pathology University of California San Diego  La Jolla California"},{"name":"Sanford Consortium for Regenerative Medicine University of California San Diego  La Jolla California"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tony","family":"Bui","sequence":"additional","affiliation":[{"name":"Department of Pathology University of California San Diego  La Jolla California"},{"name":"Sanford Consortium for Regenerative Medicine University of California San Diego  La Jolla California"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Francesca","family":"Soncin","sequence":"additional","affiliation":[{"name":"Department of Pathology University of California San Diego  La Jolla California"},{"name":"Sanford Consortium for Regenerative Medicine University of California San Diego  La Jolla California"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tao","family":"Bai","sequence":"additional","affiliation":[{"name":"Department of Neurology Northwestern University  Chicago Illinois"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"John A.","family":"Kessler","sequence":"additional","affiliation":[{"name":"Department of Neurology Northwestern University  Chicago Illinois"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mana M","family":"Parast","sequence":"additional","affiliation":[{"name":"Department of Pathology University of California San Diego  La Jolla California"},{"name":"Sanford Consortium for Regenerative Medicine University of California San Diego  La Jolla California"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4996-6792","authenticated-orcid":false,"given":"Mariko","family":"Horii","sequence":"additional","affiliation":[{"name":"Department of Pathology University of California San Diego  La Jolla California"},{"name":"Sanford Consortium for Regenerative Medicine University of California San Diego  La Jolla 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et\u00a0al. 2022. See above."},{"key":"e_1_2_13_3_3_1","unstructured":"In this paper we have shown four important points. First the initial trophoblast induction of primed human PSCs (using BMP2+IWP2) resulted in cells showing a TE\u2010like signature by transcriptome profile. Second we showed that these TE\u2010like cells can convert to TSC by applying iCTB media which resulted in a transcriptome and ELF5 promotor DNA methylation profile similar to primary TSC. Third primed PSC\u2010derived TSC can further differentiate into STB or EVT by using both in vitro lineage\u2010specific differentiation protocols and in vivo engraftment assay. Finally PSC\u2010derived TE and TSC showed similar transcriptome profile to na\u00efve (pre\u2010implantation state)\u2010PSC derived TE and TSC. These data showed that primed human ESC\/iPSC can in fact become functional TSC and further give rise to STB or EVT through lineage\u2010specific differentiation."}],"container-title":["Current Protocols"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/currentprotocols.onlinelibrary.wiley.com\/doi\/pdf\/10.1002\/cpz1.875","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,1,28]],"date-time":"2025-01-28T15:45:20Z","timestamp":1738079120000},"score":1,"resource":{"primary":{"URL":"https:\/\/currentprotocols.onlinelibrary.wiley.com\/doi\/10.1002\/cpz1.875"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2023,10]]},"references-count":51,"journal-issue":{"issue":"10","published-print":{"date-parts":[[2023,10]]}},"alternative-id":["10.1002\/cpz1.875"],"URL":"https:\/\/doi.org\/10.1002\/cpz1.875","archive":["Portico"],"relation":{},"ISSN":["2691-1299","2691-1299"],"issn-type":[{"value":"2691-1299","type":"print"},{"value":"2691-1299","type":"electronic"}],"subject":[],"published":{"date-parts":[[2023,10]]},"article-number":"e875"}}