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Publication type: Journal Article
Document type: Full Paper

Year: 1998

Author(s): Durcova-Hills, G; Prelle, K; Müller, S; Stojkovic, M; Motlik, J; Wolf, E; Brem, G

Title: Primary culture of porcine PGCs requires LIF and porcine membrane-bound stem cell factor.

Source: Zygote (6), 3 271-275.



Authors Vetmeduni Vienna:

Brem Gottfried
Müller Simone

Vetmed Research Units
Institute of Animal Breeding and Genetics


Abstract:
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, Sand 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 UF 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 SCT: 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.

Keywords Pubmed: Alkaline Phosphatase - metabolism : Animals - metabolism : Cell Culture Techniques - methods : Cell Division - drug effects : Cell Survival - drug effects : Embryo, Mammalian - metabolism : Fibroblast Growth Factor 2 - pharmacology : Germ Cells - metabolism : Growth Inhibitors - pharmacology : Growth Substances - pharmacology : Histocytochemistry - pharmacology : Humans - pharmacology : Interleukin-6 - pharmacology : Leukemia Inhibitory Factor - pharmacology : Lymphokines - pharmacology : Membrane Proteins - metabolism : Stem Cell Factor - pharmacology : Swine - pharmacology

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