Improving In Vitro Culture of Human Male Fetal Germ Cells

Male human fetal germ cells (hFGCs) give rise to spermatogonial stem cells (SSCs), which are the adult precursors of the male gametes. Human SSCs are a promising (autologous) source of cells for male fertility preservation; however, in contrast to mouse SSCs, we are still unable to culture them in t...

Full description

Saved in:
Bibliographic Details
Published in:Cells (Basel, Switzerland) Switzerland), 2021-08, Vol.10 (8), p.2033
Main Authors: Martin-Inaraja, Myriam, Ferreira, Monica, Taelman, Jasin, Eguizabal, Cristina, Chuva De Sousa Lopes, Susana M.
Format: Article
Language:English
Subjects:
Antibodies
Autografts
Cell culture
Culture media
Embryos
extracellular matrix substrate
Fetuses
Fibroblast growth factor 2
Fibroblasts
Gametes
Gametogenesis
Gelatin
Germ cells
Glial cell line-derived neurotrophic factor
Growth factors
human
Hydrogels
Laminin
male fetal germ cells
Monkeys & apes
Oct-4 protein
Spermatogenesis
Stem cell transplantation
Stem cells
tissue culture
Vitronectin
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:Male human fetal germ cells (hFGCs) give rise to spermatogonial stem cells (SSCs), which are the adult precursors of the male gametes. Human SSCs are a promising (autologous) source of cells for male fertility preservation; however, in contrast to mouse SSCs, we are still unable to culture them in the long term. Here, we investigated the effect of two different culture media and four substrates (laminin, gelatin, vitronectin and matrigel) in the culture of dissociated second trimester testes, enriched for hFGCs. After 6 days in culture, we quantified the presence of POU5F1 and DDX4 expressing hFGCs. We observed a pronounced difference in hFGC number in different substrates. The combination of gelatin-coated substrate and medium containing GDNF, LIF, FGF2 and EGF resulted in the highest percentage of hFGCs (10% of the total gonadal cells) after 6 days of culture. However, the vitronectin-coated substrate resulted in a comparable percentage of hFGCs regardless of the media used (3.3% of total cells in Zhou-medium and 4.8% of total cells in Shinohara-medium). We provide evidence that not only the choices of culture medium but also choices of the adequate substrate are crucial for optimizing culture protocols for male hFGCs. Optimizing culture conditions in order to improve the expansion of hFGCs will benefit the development of gametogenesis assays in vitro.
ISSN:2073-4409
2073-4409
DOI:10.3390/cells10082033