Primary culture of porcine PGCs requires LIF and porcine membrane-bound stem cell factor.
We studied the effect of murine leukaemia inhibitory factor (LIF), human basic fibroblast growth factor (bFGF) and porcine stem cell factor (SCF) on the survival and/or proliferation of porcine primordial germ cells (PGCs) obtained from 27-day-old embryos in vitro. PGCs were cultured in embryonic stem cell (ESC) medium supplemented with or without either LIF (1000 IU/ml) alone or LIF together with bFGF (10 ng/ml). They were seeded on mitotically inactivated feeder cells, either STO or transfected STO cells (STO#8), expressing the membrane-bound form of porcine SCF. PGCs were identified by their alkaline phosphatase (AP) activity and counted after 1, 3 and 5 days in culture. After 1 day of culture, PGCs cultured on STO#8 cells showed significantly higher survival than PGCs cultured on STO cells (p < 0.05). The combined effect of SCF and LIF caused a significant increase in PGC number by day 3 of culture when PGCs were cultured on either STO cells (p < 0.01) or STO#8 (p < 0.001). When SCF and LIF were used together with bFGF no increase in the PGC number was observed. Our results suggest that the membrane-bound form of porcine SCF plays a pivotal role in the primary culture of porcine PGCs and that bFGF is not required in vitro.
Durcova-Hills
G
G
Institute of Animal Physiology and Genetics, Libechov, Czech Republic.
Prelle
K
K
Müller
S
S
Stojkovic
M
M
Motlik
J
J
Wolf
E
E
Brem
G
G
D000469:Alkaline Phosphatase / Q000378:metabolism; D000818:Animals; D018929:Cell Culture Techniques / Q000379:methods; D002455:Cell Division / Q000187:drug effects*; D002470:Cell Survival / Q000187:drug effects; D004622:Embryo, Mammalian / Q000378:metabolism; D016222:Fibroblast Growth Factor 2 / Q000494:pharmacology; D005854:Germ Cells / Q000378:metabolism*; D006131:Growth Inhibitors / Q000494:pharmacology*; D006133:Growth Substances / Q000494:pharmacology; D006651:Histocytochemistry; D006801:Humans; D015850:Interleukin-6; D053605:Leukemia Inhibitory Factor; D008222:Lymphokines / Q000494:pharmacology*; D008565:Membrane Proteins / Q000378:metabolism; D019089:Stem Cell Factor / Q000494:pharmacology*; D013552:Swine
D006131:Growth Inhibitors; D006133:Growth Substances; D015850:Interleukin-6; C507837:LIF protein, human; D053605:Leukemia Inhibitory Factor; D008222:Lymphokines; D008565:Membrane Proteins; D019089:Stem Cell Factor; D016222:Fibroblast Growth Factor 2; D000469:Alkaline Phosphatase