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

Published online by Cambridge University Press:  01 August 1998

Gabriela Durcova-Hills
Affiliation:
Institute of Animal Physiology and Genetics, Libechov, Czech Republic Institute of Animal Breeding and Genetics, Vienna, Austria
Katja Prelle
Affiliation:
Department of Molecular Animal Breeding and Genetics, Oberschleissheim, Germany
Sigrid Müller
Affiliation:
Institute of Animal Breeding and Genetics, Vienna, Austria
Miodrag Stojkovic
Affiliation:
Department of Molecular Animal Breeding and Genetics, Oberschleissheim, Germany
Jan Motlik
Affiliation:
Institute of Animal Physiology and Genetics, Libechov, Czech Republic
Eckhard Wolf
Affiliation:
Department of Molecular Animal Breeding and Genetics, Oberschleissheim, Germany
Gottfried Brem
Affiliation:
Institute of Animal Breeding and Genetics, Vienna, Austria

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, 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.

Type
Research Article
Copyright
© 1998 Cambridge University Press

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