Propagation of goat putative spermatogonial stem cells under growth factors defined serum-free culture conditions

Culture of spermatogonial stem cells and use of surrogate sires as a breeding technology to propagate superior genetics in livestock production: A systematic review

“…The long-term culture of SSCs involved either of the two strategies: First, by supplementation of culture medium with a cocktail of growth factors (GDNF, bFGF, LIF, stem cell factor, and SDF); second, by growing SSCs on a feeder cell layer or feeder cellsfree culture using preconditioned serum-free media. Early studies utilized serum as one of the important components of culture medium, although published reports have shown that the presence of serum in the medium enhances the growth of somatic cells and inhibits SSC self-renewal [3,5,6,21]. Culture studies of goat SSC in media containing 10% FBS have been documented [14,15].…”

“…Additionally, transplantation of the cultured SSC which is a definitive test to verify the stem cell capacity of cultured SSC was not carried out. Recent studies for goat SSC have demonstrated the detrimental effects of serum on SSC self-renewal and the enhanced proliferation of somatic cells in SSC culture medium containing serum [21,29]. The long-term culture of SSC has failed mainly due to difficulties in providing all in vitro conditions that mimic the in vivo SSC niche, which has physical, mechanical, and chemical support by the surrounding somatic cells and lack of unique markers for SSC identification in culture [90].…”

“…Evaluation of SSC expression of the LIN28 gene was conducted in only one study and was suggested to be uniquely expressed by SSC [3]. From the review, different studies used different markers for the verification of SSC undifferentiated status; however, expression of PLZF as being confined to SSC only not only rodents but also livestock has been supported by recent studies in bovine [3,5], sheep [17,18], pig [19], and goat [20,21].…”

Evaluation of Porcine Somatic and Testicular Germ Cells During Differentiation and Proliferation

“…The prepared slides were divided into two groups and incubated with either primary sox9 antibody or vimentin antibody (Sigma) and speciesspeci c secondary antibodies. The characterization of labelled cells was analyzed by confocal laser scanning microscope Zeiss LSM (Zeiss, Germany) [20].…”

“…The labelled cells were identi ed by simple nuclear counterstain with 0.2 µg/ml 4', 6-diamidino-2-phenylindole (DAPI) dye. Labelled positive cells with antibodies were considered by a confocal laser scanning microscope Zeiss LSM 700, and images of cells were obtained using a Zeiss LSM-TPMT camera [20,23,24].…”