AUTHOR=Nabulindo Nakami Wilkister , Nguhiu-Mwangi James , Kipyegon Ambrose Ng'eno , Ogugo Moses , Muteti Charity , Christian Tiambo , Oatley Melissa J. , Oatley Jon M. , Kemp Stephen 

TITLE=Culture of Kenyan Goat (Capra hircus) Undifferentiated Spermatogonia in Feeder-Free Conditions

JOURNAL=Frontiers in Veterinary Science

VOLUME=Volume 9 - 2022

YEAR=2022

URL=https://www.frontiersin.org/journals/veterinary-science/articles/10.3389/fvets.2022.894075

DOI=10.3389/fvets.2022.894075

ISSN=2297-1769

ABSTRACT=The undifferentiated spermatogonial population in mammalian testes contains a spermatogenic stem cell (SSC) population that can regenerate continual spermatogenesis following transplantation. This capacity has the potential to be exploited as a surrogate sires breeding tool to achieve widespread dissemination of desirable genetics in livestock production. Because SSCs are relatively rare in testicular tissue, the ability to expand a population in vitro would be advantageous to provide large numbers for transplantation into surrogate recipient males. Here, we evaluated conditions that would support long-term in vitro maintenance of undifferentiated spermatogonia from a goat breed that is endemic to Kenyan livestock production. Single-cell suspensions enriched for undifferentiated spermatogonia from prepubertal bucks were seeded on laminin-coated tissue culture plates and maintained in Stempro serum-free medium that had been conditioned by goat fetal fibroblasts and supplemented with a growth factor cocktail that included GDNF, LIF, SDF, and FGF. Over a 45-day period, the primary cultures developed a cluster morphology indicative of in vitro grown undifferentiated spermatogonia from other species and expressed the tell-tale germ cell marker VASA as well as the previously defined spermatogonial marker PLZF. Taken together, these findings provide a methodology for isolating the SSC containing undifferentiated spermatogonial population from goat testes and long-term maintenance in defined culture conditions.