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

Zygote. 1998 Aug;6(3):271-5. doi: 10.1017/s0967199498000215.

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Alkaline Phosphatase / metabolism
  • Animals
  • 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
  • Humans
  • Interleukin-6*
  • Leukemia Inhibitory Factor
  • Lymphokines / pharmacology*
  • Membrane Proteins / metabolism
  • Stem Cell Factor / pharmacology*
  • Swine

Substances

  • Growth Inhibitors
  • Growth Substances
  • Interleukin-6
  • LIF protein, human
  • Leukemia Inhibitory Factor
  • Lymphokines
  • Membrane Proteins
  • Stem Cell Factor
  • Fibroblast Growth Factor 2
  • Alkaline Phosphatase