Abstract
Primary human trophoblast stem cells (TSCs) and TSCs derived from human pluripotent stem cells (hPSCs) can potentially model placental processes in vitro. Yet, the pluripotent states and factors involved in the differentiation of hPSCs to TSCs remain poorly understood. In this study, we demonstrate that the primed pluripotent state can generate TSCs by activating pathways such as Epidermal Growth Factor (EGF) and Wingless-related integration site (WNT), and by suppressing tumor growth factor beta (TGFβ), histone deacetylases (HDAC), and Rho-associated protein kinase (ROCK) signaling pathways, all without the addition of exogenous Bone morphogenetic protein 4 (BMP4)—a condition we refer to as the TS condition. We characterized this process using temporal single-cell RNA sequencing to compare TS conditions with differentiation protocols involving BMP4 activation alone or BMP4 activation in conjunction with WNT inhibition. The TS condition consistently produced a stable, proliferative cell type that closely mimics first-trimester placental cytotrophoblasts, marked by the activation of endogenous retroviral genes and the absence of amnion expression. This was observed across multiple cell lines, including various primed induced pluripotent stem cell (iPSC) and embryonic stem cell (ESC) lines. Primed-derived TSCs can proliferate for over 30 passages and further specify into multinucleated syncytiotrophoblasts and extravillous trophoblast cells. Our research establishes that the differentiation of primed hPSCs to TSC under TS conditions triggers the induction of TMSB4X, BMP5/7, GATA3, and TFAP2A without progressing through a naive state. These findings propose that the primed hPSC state is part of a continuum of potency with the capacity to differentiate into TSCs through multiple routes.
| Original language | English (US) |
|---|---|
| Article number | 3291 |
| Journal | Scientific reports |
| Volume | 14 |
| Issue number | 1 |
| DOIs | |
| State | Published - Dec 2024 |
Funding
Supported by funding from the Lieber Institute for Brain Development and a NARSAD Young Investigator Grant from the Brain & Behavior Research Foundation to JAE, the National Institute of Health (NIH) T32 Fellowship (T32MH015330) to KJB. The authors are grateful for the financial support of the Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)—Financing code 001 (ARB, VE and AF). This work was also supported in part by FAPESP—Sao Paulo Research Foundation ( http://www.fapesp.br/en/ ): Grant Number 2018/18560-6. ChatGPT was used to correct grammatical errors and improve text flow. We thank Bill Ulrich for the scRNA-seq browser. We thank Tricia Nilles and Worod Allak from the Becton Dickinson Immune Function Laboratory at the Johns Hopkins Bloomberg School of Public Health, for flow cytometry technical assistance. The facility was supported in part by CFAR: P30AI094189-04 (Chaisson). Supported by funding from the Lieber Institute for Brain Development and a NARSAD Young Investigator Grant from the Brain & Behavior Research Foundation to JAE, the National Institute of Health (NIH) T32 Fellowship (T32MH015330) to KJB. The authors are grateful for the financial support of the Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)—Financing code 001 (ARB, VE and AF). This work was also supported in part by FAPESP—Sao Paulo Research Foundation (http://www.fapesp.br/en/): Grant Number 2018/18560-6. ChatGPT was used to correct grammatical errors and improve text flow. We thank Bill Ulrich for the scRNA-seq browser. We thank Tricia Nilles and Worod Allak from the Becton Dickinson Immune Function Laboratory at the Johns Hopkins Bloomberg School of Public Health, for flow cytometry technical assistance. The facility was supported in part by CFAR: P30AI094189-04 (Chaisson).
ASJC Scopus subject areas
- General