Differentiation and Expansion of Human Extra-Embryonic Endoderm Cell Lines from Naïve Pluripotent Stem Cells
Research output: Chapter in Book/Report/Conference proceeding › Book chapter › Research › peer-review
In human, endoderm is induced in two waves, with the first being the extra-embryonic primitive endoderm (PrE), otherwise known as hypoblast, induced during blastocyst development, and the second being gastrulation-stage definitive endoderm (DE). The PrE gives rise to the primary and secondary yolk sac, and has supportive functions during pregnancy for nutrient provision, with descendants of this extra-embryonic lineage also playing a role in embryonic patterning. As in DE specification, we recently found that PrE could be induced in vitro by Wnt and Nodal-related signaling, but that the critical difference was in the pluripotent starting point for differentiation. Thus, blastocyst-like naïve human pluripotent stem cells retain the unique capacity to differentiate into PrE cultures, a cell type resembling the pre-implantation hypoblast. The PrE cells could then be expanded as stable naïve extra-embryonic endoderm (nEnd) cell lines, capable of indefinite self-renewal. Here, we describe detailed protocols to differentiate naïve pluripotent stem cells into PrE and then expand the cultures as nEnd, including descriptions of morphology, passaging technique, and troubleshooting.
Original language | English |
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Title of host publication | Methods in Molecular Biology |
Number of pages | 12 |
Publisher | Humana Press |
Publication date | 2022 |
Pages | 105-116 |
ISBN (Print) | 978-1-0716-1910-0 |
ISBN (Electronic) | 978-1-0716-1908-7 |
DOIs | |
Publication status | Published - 2022 |
Series | Methods in Molecular Biology |
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Volume | 2416 |
ISSN | 1064-3745 |
Bibliographical note
Publisher Copyright:
© 2022, The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.
- Embryonic stem cells, Endoderm, Extra-embryonic, Human, Hypoblast, Naïve, Pluripotency, Primitive endoderm
Research areas
ID: 342675337