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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 ,
Katja Prelle ,
Sigrid Müller ,
Miodrag Stojkovic ,
Jan Motlik ,
Eckhard Wolf  and
Gottfried Brem
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
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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.

Keywords

Cell proliferationGrowth factorsPorcine primordial germ cells
Type
Research Article
Information
Zygote , Volume 6 , Issue 3 , August 1998 , pp. 271 - 275
Copyright
© 1998 Cambridge University Press

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