Culture supplementation of bFGF, GDNF, and LIF alters in vitro proliferation, colony formation, and pluripotency of neonatal porcine germ cells

Effects of Growth Factors on In Vitro Culture of Neonatal Piglet Testicular Tissue Fragments

“…These findings suggest that bFGF also has a direct or indirect role in the mitogenic and proliferative activity of germ cells and/or somatic cells [80]. Similar to our findings, bFGF supplementation increased the number of early germ cells in mice [24], bovine [78], pigs [5], and humans [81]. EGF has also been described as a spermatogonial stimulating growth factor [82]; however, in the present study, we observed that the EGFsupplemented group had the lowest number of early germ cells.…”

“…The present study was designed to examine the in vitro effects of various growth factors on piglet testicular tissue fragments. The concentrations of growth factors used in the present study were based on our previous in vivo and in vitro studies on pig testis cells and tissue [ 5 , 40 , 41 ], or relevant other work [ 7 ]. As schematically shown in Figure 1 , testes collected from 1-wk-old piglets were divided into small testicular fragments and cultured in 6 differently supplemented media for 8 weeks where samples were taken every 2 weeks.…”

“…The long-term culture of rodent SSCs is relatively well established [ 2 ], while this knowledge is largely lacking in domestic animals [ 3 ]. However, the short-term culture of SSCs has been reported in different farm animals, including pigs [ 4 , 5 , 6 ], goats [ 7 , 8 , 9 , 10 ], buffalo [ 11 , 12 , 13 ], and sheep [ 14 ].…”

“…GDNF has also been reported to increase the number of SSCs in buffalo [ 13 ] and goats [ 10 ] during in vitro culturing. Recently, we observed that GDNF increases porcine gonocyte/SSC proliferation in a dose-dependent manner in a two-dimensional (2D) cell culture [ 5 ]. It has been shown that GDNF promotes the proliferation of gonocytes/SSCs via activation of the Src kinase(s) and a PI3K/Akt pathway to increase N-myc gene expression [ 25 ].…”

“…However, we recently reported that bFGF alone or combined with GDNF could increase the number of porcine gonocytes/SSCs in a culture. Interestingly, after 28 days of gonocyte culturing, groups treated with bFGF alone or in combination with GDNF did not differ, suggesting that bFGF alone is also sufficient for the long-term culture of porcine gonocytes [ 5 ].…”