Embryonic germ cells (EG cells) are pluripotential undifferentiated stem cells isolated from cultured primordial germ cells (PGCs). Like ES cells, EG cells are of importance for gene targeting, therapeutical cloning a...Embryonic germ cells (EG cells) are pluripotential undifferentiated stem cells isolated from cultured primordial germ cells (PGCs). Like ES cells, EG cells are of importance for gene targeting, therapeutical cloning and organ transplantation. The aim of this study was to isolate and characterize EG cells from porcine PGCs. The genital ridges from 24-26 days old porcine embryos were treated in 0 02% EDTA for 20 min and pricked with a needle to release PGCs. The isolated PGCs were cultured on a SNL feeder layer in an EG cell medium. The EG cell medium consisted of Dulbecco’s modified Eagle’s medium (DMEM) supplemented with 20% Buffalo rat lever (BRL) cell-conditioned medium, 15% fetal bovine serum, 1 mmol L-glutamine, 0 1 mol nonessential amino acids, 10 μmol β-mercaptoethanol and antibiotics. The freshly isolated PGCs were positive for alkaline phosphatase activity and Periodic acid-Schiff’s staining. Under this culture regime, PGCs could be maintained in an undifferentiated state and used for further cultures. One strain of the cultured PGCs was cultured 8 times, and alkaline phosphatase activity was detected in the colony formed from this strain. These cultured PGCs could spontaneously differentiate into fibroblast-like cells. These data suggested that we had successfully isolated EG-like cells from porcine PGCs .展开更多
文摘Embryonic germ cells (EG cells) are pluripotential undifferentiated stem cells isolated from cultured primordial germ cells (PGCs). Like ES cells, EG cells are of importance for gene targeting, therapeutical cloning and organ transplantation. The aim of this study was to isolate and characterize EG cells from porcine PGCs. The genital ridges from 24-26 days old porcine embryos were treated in 0 02% EDTA for 20 min and pricked with a needle to release PGCs. The isolated PGCs were cultured on a SNL feeder layer in an EG cell medium. The EG cell medium consisted of Dulbecco’s modified Eagle’s medium (DMEM) supplemented with 20% Buffalo rat lever (BRL) cell-conditioned medium, 15% fetal bovine serum, 1 mmol L-glutamine, 0 1 mol nonessential amino acids, 10 μmol β-mercaptoethanol and antibiotics. The freshly isolated PGCs were positive for alkaline phosphatase activity and Periodic acid-Schiff’s staining. Under this culture regime, PGCs could be maintained in an undifferentiated state and used for further cultures. One strain of the cultured PGCs was cultured 8 times, and alkaline phosphatase activity was detected in the colony formed from this strain. These cultured PGCs could spontaneously differentiate into fibroblast-like cells. These data suggested that we had successfully isolated EG-like cells from porcine PGCs .
