Optimization of the conditions of isolation and culture of dairy goat male germline stem cells (mGSC)

Multipotent male germline stem cells (mGSCs) from neonate porcine testis

“…In our study, we also found that the efficiency of establishment of pmGSC line is greatly affected by the age of porcine. This is consistent with previously established in the mouse and dairy goat (Zhu et al 2012). More important, the growth curve and cell cycle analysis demonstrated that porcine mGSCs cultured in the serum-and feeder-free system could be expanded and maintained stably in the presence of N2B27 and supplementation of growth factors for two month.…”

“…Niche and microenvironments play important roles in the fate and behavior of stem cells (Griswold et al 2013;DeFalco et al 2015). Recent studies demonstrate the importance of growth factors and extracellular matrix (ECM) on stem cell and germ cell behavior (Kuijk et al 2009;Zhu et al 2012). Derivation of pluripotent cells from male germ cells is enhanced by the cytokines, such as epidermal growth factor (EGF), Colony stimulating factor 1 (CSF1), Leukemia inhibitory factor (LIF), Glial cell-derived neurotrophic factor (GDNF) (Kuijk et al 2009;Zhu et al 2012).…”

“…Recent studies demonstrate the importance of growth factors and extracellular matrix (ECM) on stem cell and germ cell behavior (Kuijk et al 2009;Zhu et al 2012). Derivation of pluripotent cells from male germ cells is enhanced by the cytokines, such as epidermal growth factor (EGF), Colony stimulating factor 1 (CSF1), Leukemia inhibitory factor (LIF), Glial cell-derived neurotrophic factor (GDNF) (Kuijk et al 2009;Zhu et al 2012). GDNF is secreted by the somatic Sertoli cells and peritubular cells, and is a well-known inducer of mouse SSC proliferation in vitro (Oatley et al 2008).…”

“…All procedures were approved by Shaanxi Centre of Stem Cells Engineering & Technology, Northwest A&F University. The porcine were killed by standard methods and porcine testicular cells were isolated in our laboratory as previous report (Zhu et al 2012), using CDD mixed digestive enzyme: 2.0 mg/mL collagenase IV (Invitrogen) + 20 μg/mL DNase (Roche) + 2.0 mg/mL dispase (Invitrogen). The dissociated cells were suspended in DMEM (Invitrogen) containing 10% FBS (Hyclone, Logan, UT), 2.0 mM Lglutamine (Invitrogen), 1% nonessential amino acids (Invitrogen), 0.1 mM β-mercaptoethanol (Sigma), 100 unit/mL penicillin and 100 mg/mL streptomycin (Invitrogen).…”

Spermatogonial stem cells: Current biotechnological advances in reproduction and regenerative medicine

“…Culture conditions for SSC propagation in vitro were first developed in mice [16,20] . This was the base for the development of domestic animals SSC culture systems [7][8][9][10][21][22][23][24][25] . Most of these systems have in common the use of GDNF and serum.…”