ABSTRACT
Objective To test whether feeder cells derived from mouse embryonic stem cells(mESCs) could support the growth of mESCs themselves. Methods mESCs were induced to form mouse embryoid bodies(EB),and then fibroblast-like cells were derived from further differentiated mEB(mEB-dF),which served as feeder cells.The undifferentiation of mESCs grown on mEB-dF was confirmed by morphological analysis,colony efficiency and cell differentiation rate of mESCs,immunocytochemistry,alkaline phosphatase staining and RT-PCR.The pluripotency of mESCs grown on mEB-dF was examined by RT-PCR,inducing their differentiation in vivo and in vitro. Results(Forty-eight) fibroblast-like cells lines were derived from the same EB at three periods(d 10,d 15 and d 20),and five of them,mostly derived from d15 EB,were able to maintain mESCs in undifferentiated status and pluripotential ability over 10 passages.mESCs cultured on these feeder cell lines expressed alkaline phosphatase and specific mESCs markers,including SSEA-1,OCT-4,NANOG,and formed EB in vitro and teratomas in vivo.However,the majority of mEB-dF lines(43/48) has no such ability. Conclusion This study not only provides a novel feeder system for mESCs culture,avoiding lot of disadvantages of mouse embryo fibroblasts used as the feeder,but also indicates that fibroblast-like cells derived from mESCs take on different functions.The molecular mechanism of different function of these fibroblast cells is worthy of further investigations.
ABSTRACT
To investigate the differences of proliferation capacity and phenotype properties of mesenchymal stem cells (MSCs) derived from bone marrow (BM) of aplastic anemia patients, fetuses and children, MSCs were isolated from BM of patients with aplastic anemia and expanded in vitro; MSCs derived from BM of fetuses and children were used as normal control groups, three sources of MSCs were compared by morphology, in vitro proliferation capacity, phenotype and immunocytochemistry. The results showed that MSCs could be isolated and expanded from aplastic anemia patient BM. MSCs derived from BM of aplastic anemia patients shared a similar morphology and phenotype with derived MSCs from BM of fetuses and children. However, in vitro proliferation capacity of MSCs derived from BM of aplastic anemia patients after 20 population doublings (PD) was significantly lower, compared with MSCs from BM of fetuses and children. BM MSCs derived from children and fetuses proliferated for more than 30 PD. It is concluded that BM MSCs from aplastic anemia patients appears to be normal in phenotype but their proliferation capacity is lower in comparison with control groups.