Efficient generation of marmoset primordial germ cell-like cells using induced pluripotent stem cells
- University of Pennsylvania, United States
- The University of Texas at San Antonio, United States
Abstract
Reconstitution of germ cell fate from pluripotent stem cells provides an opportunity to understand the molecular underpinnings of germ cell development. Here, we established robust methods for pluripotent stem cell (iPSC) culture in the common marmoset (Callithrix jacchus, cj), allowing stable propagation in an undifferentiated state. Notably, iPSCs cultured on a feeder layer in the presence of a WNT signaling inhibitor upregulated genes related to ubiquitin-dependent protein catabolic processes and enter a permissive state that enables differentiation into primordial germ cell-like cells (PGCLCs) bearing immunophenotypic and transcriptomic similarities to pre-migratory cjPGCs in vivo. Induction of cjPGCLCs is accompanied by transient upregulation of mesodermal genes, culminating in the establishment of a primate specific germline transcriptional network. Moreover, cjPGCLCs can be expanded in monolayer while retaining the germline state. Upon co-culture with mouse testicular somatic cells, these cells acquire an early prospermatogonia-like phenotype. Our findings provide a framework for understanding and reconstituting marmoset germ cell development in vitro, thus providing a comparative tool and foundation for a preclinical modeling of human in vitro gametogenesis.
Data availability
Accession number for RNA-seq and whole genome bisulfite sequencing generated in this study is GSE210576. The Exome sequence data generated in this study are available as SRA BioProject PRJNA856282.
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Efficient generation of marmoset primordial germ cell-like cells using induced pluripotent stem cellsNCBI Gene Expression Omnibus, GSE210576.
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Sequencing of Callithrix jacchus iPSC linesSRA BioProject, PRJNA856282.
Article and author information
Author details
Funding
Open Philanthropy Project (2019-197906)
- Kotaro Sasaki
Open Philanthropy Project (10080664)
- Kotaro Sasaki
National Institute on Drug Abuse (U01DA054170)
- Brian P Hermann
Eunice Kennedy Shriver National Institute of Child Health and Human Development (R01HD090007)
- Brian P Hermann
National Institute on Aging (P51OD011133)
- Corinna N Ross
National Institute on Minority Health and Health Disparities (G12MD007591)
- Sean Vargas
National Science Foundation (DBI-1337513)
- Sean Vargas
National Science Foundation (DBI-2018408)
- Sean Vargas
The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.
Ethics
Animal experimentation: This study was performed in strict accordance with the recommendations in the Guide for the Care and Use of Laboratory Animals of the National Institutes of Health. All procedures were reviewed and approved by the Texas Biomedical Research Institute IACUC (1772CJ).
Copyright
© 2023, Seita et al.
This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.
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Efficient generation of marmoset primordial germ cell-like cells using induced pluripotent stem cells
eLife 12:e82263.
https://doi.org/10.7554/eLife.82263
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