원문정보
초록
영어
Germ cells are alternative cell sources for deriving pluripotent stem cells. When cultured with feeder cells and adequate cytokines, migrating primordial germ cells (PGCs) can be reprogrammed into pluripotent stem cells, named embryonic germ cells (EGCs). In this study, we attempted to establish and characterize pig embryonic germ cells from fetal gonads. Consequently, EGC lines were derived from the genital ridges of a porcine dpc 30 fetuses in media containing LIF, FGF2 and SCF. After establishment, this cells were cultured and stabilized in LIF or FGF2 contained media. These cell lines were maintained in both condition over an extended time period and were able to spontaneously differentiate into the three germ layers in vitro. Interestingly, cell lines cultured in LIF or FGF2 expressed different pluripotency markers. While LIF-treated pig EGCs (LIF-pEGCs) expressed only few pluripotent markers including OCT4, SOX2 and NANOG, FGF2-treated pEGCs (FGF2-pEGCs) expressed pluripotency markers such as OCT4, SOX2, NANOG and SSEA4. As a result of molecular analysis, it was verified that LIF- and FGF2-EGCs represented intermediated state of EGCs and complete reprogrammed EGCs respectively, and reprogramming of pig gonadal PGCs is progressed in stepwise manner, from PGCs via intermediated EGCs to EGCs. In conclusion, it is verified that FGF2 signaling have important roles in reprogramming and maintaining pEGCs from fetal gonads.