Derivation of pluripotential embryonic stem cells from murine primordial germ cells in culture.
Steel factor (SF) and LIF (leukemia inhibitory factor) synergistically promote the proliferation and survival of mouse primordial germ cells (PGCs), but only for a limited time period in culture. We show here that addition of bFGF to cultures in the presence of membrane-associated SF and LIF enhances the growth of PGCs and allows their continued proliferation beyond the time when they normally stop dividing in vivo. They form colonies of densely packed, alkaline phosphatase-positive, SSEA-1-positive cells resembling undifferentiated embryonic stem (ES) cells in morphology. These cultures can be maintained on feeder layers for at least 20 passages, and under appropriate conditions give rise to embryoid bodies and to multiple differentiated cell phenotypes in monolayer culture and in tumors in nude mice. PGC-derived ES cells can also contribute to chimeras when injected into host blastocysts. The long-term culture of PGCs and their reprogramming to pluripotential ES cells has important implications for germ cell biology and the induction of teratocarcinomas.
Matsui
Y
Y
Department of Cell Biology Vanderbilt University Medical School Nashville, Tennessee 37232.
Zsebo
K
K
Hogan
B L
BL
D000469:Alkaline Phosphatase / Q000032:analysis / Q000378:metabolism; D000818:Animals; D002455:Cell Division / Q000187:drug effects; D002478:Cells, Cultured / Q000187:drug effects; D016222:Fibroblast Growth Factor 2 / Q000494:pharmacology; D006131:Growth Inhibitors / Q000494:pharmacology*; D016298:Hematopoietic Cell Growth Factors / Q000494:pharmacology*; D015850:Interleukin-6; D053605:Leukemia Inhibitory Factor; D008222:Lymphokines / Q000494:pharmacology*; D051379:Mice; D008815:Mice, Inbred Strains; D019089:Stem Cell Factor; D013234:Stem Cells / Q000187:drug effects*
D006131:Growth Inhibitors; D016298:Hematopoietic Cell Growth Factors; D015850:Interleukin-6; D053605:Leukemia Inhibitory Factor; C507838:Lif protein, mouse; D008222:Lymphokines; D019089:Stem Cell Factor; D016222:Fibroblast Growth Factor 2; D000469:Alkaline Phosphatase