Two distinct trophectoderm lineage stem cells from human pluripotent stem cells

Adam Mischler,Victoria Karakis,Jessica Mahinthakumar,Celeste K Carberry,Adriana San Miguel,Julia E Rager,Rebecca C Fry,Balaji M Rao

doi:10.1016/j.jbc.2021.100386

Adam Mischler, Victoria Karakis + Show 6 more

Open Access

https://doi.org/10.1016/j.jbc.2021.100386

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Abstract

The trophectoderm layer of the blastocyst-stage embryo is the precursor for all trophoblast cells in the placenta. Human trophoblast stem (TS) cells have emerged as an attractive tool for studies on early trophoblast development. However, the use of TS cell models is constrained by the limited genetic diversity of existing TS cell lines and restrictions on using human fetal tissue or embryos needed to generate additional lines. Here we report the derivation of two distinct stem cell types of the trophectoderm lineage from human pluripotent stem cells. Analogous to villous cytotrophoblasts in vivo, the first is a CDX2- stem cell comparable with placenta-derived TS cells—they both exhibit identical expression of key markers, are maintained in culture and differentiate under similar conditions, and share high transcriptome similarity. The second is a CDX2+ stem cell with distinct cell culture requirements, and differences in gene expression and differentiation, relative to CDX2- stem cells. Derivation of TS cells from pluripotent stem cells will significantly enable construction of in vitro models for normal and pathological placental development.

Highlights

The trophectoderm layer of the blastocyst-stage embryo is the precursor for all trophoblast cells in the placenta
Media formulations in previous studies on trophoblast differentiation of human embryonic stem cell (hESC) included components such as knockout serum replacement (KSR) or bovine serum albumin (BSA) that act as carriers for lipids
We have shown that two distinct stem cell populations of the trophectoderm lineage, human pluripotent stem cell (hPSC)-trophoblast stem (TS) and hPSC-TSCDX2, can be derived from hESCs and human induced pluripotent stem cell (hiPSC) under chemically defined culture conditions

Summary

Introduction

The trophectoderm layer of the blastocyst-stage embryo is the precursor for all trophoblast cells in the placenta. The trophectoderm forms the cytotrophoblast (CTB), a putative stem cell that can differentiate to form the two major cell types in the placenta, the extravillous trophoblast (EVT) and the syncytiotrophoblast (STB) [1, 2]. Trophoblast stem (TS) cells derived from first-trimester human placental samples and blastocyst-stage embryos have emerged as an attractive in vitro model system for early human trophoblast [5]. Unlike early gestation primary samples where the projected pregnancy outcome is uncertain, human induced pluripotent stem cells (hiPSCs) can potentially provide models of validated normal and pathological trophoblast development [6]. We report the derivation and maintenance of two distinct trophectoderm lineage stem cell types from hPSCs, human embryonic stem cells (hESCs) and hiPSCs, in chemically defined culture conditions. The routine derivation of TS cells from hPSCs will provide powerful tools for mechanistic studies on normal and pathological early trophoblast development

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