ArticlePloS one2014
Three-step method for proliferation and differentiation of human embryonic stem cell (hESC)-derived male germ cells.
Article in PloS one, 2014. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 10 papers.
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Who cites it
10 citing papers in PubMed, 26 citations in OpenAlex.
- Establishment and optimization of the two-step induction system for generating primordial germ cell-like cells from chicken embryonic stem cells.FEBS open bio · 2026Article
- Artificial Gametogenesis and In Vitro Spermatogenesis: Emerging Strategies for the Treatment of Male Infertility.International journal of molecular sciences · 2025Review
- Testicular immunity.Molecular aspects of medicine · 2024Review
- The Evolutionary Route of in vitro Human Spermatogenesis: What is the Next Destination?Stem cell reviews and reports · 2024Review
- Comprehensive chromatin proteomics resolves functional phases of pluripotency and identifies changes in regulatory components.Nucleic acids research · 2023Article
- Adaptation of Human Testicular Niche Cells for Pluripotent Stem Cell and Testis Development Research.Tissue engineering and regenerative medicine · 2020Article
- Human Pluripotent Stem Cells in Reproductive Science-a Comparison of Protocols Used to Generate and Define Male Germ Cells from Pluripotent Stem Cells.International journal of molecular sciences · 2020Review
- Stem Cells as a Resource for Treatment of Infertility-related Diseases.Current molecular medicine · 2019Review
- CHIR99021 combined with retinoic acid promotes the differentiation of primordial germ cells from human embryonic stem cells.Oncotarget · 2017Article
- Induction of Germ Cell-like Cells from Porcine Induced Pluripotent Stem Cells.Scientific reports · 2016Article
Corrections and comments
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Authors and funding
4 authors at 2 institutions in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
The low efficiency of differentiation into male germ cell (GC)-like cells and haploid germ cells from human embryonic stem cells (hESCs) reflects the culture method employed in the two-dimensional (2D)-microenvironment. In this study, we applied a three-step media and calcium alginate-based 3D-culture system for enhancing the differentiation of hESCs into male germ stem cell (GSC)-like cells and haploid germ cells. In the first step, embryoid bodies (EBs) were derived from hESCs cultured in EB medium for 3 days and re-cultured for 4 additional days in EB medium with BMP4 and RA to specify GSC-like cells. In the second step, the resultant cells were cultured in GC-proliferation medium for 7 days. The GSC-like cells were then propagated after selection using GFR-α1 and were further cultured in GC-proliferation medium for 3 weeks. In the final step, a 3D-co-culture system using calcium alginate encapsulation and testicular somatic cells was applied to induce differentiation into haploid germ cells, and a culture containing approximately 3% male haploid germ cells was obtained after 2 weeks of culture. These results demonstrated that this culture system could be used to efficiently induce GSC-like cells in an EB population and to promote the differentiation of ESCs into haploid male germ cells.
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