title: Human fetal liver stromal cells that overexpress bFGF support growth and maintenance of human embryonic stem cells.
abstract: In guiding hES cell technology toward the clinic, one key issue to be addressed is to culture and maintain hES cells much more safely and economically in large scale. In order to avoid using mouse embryonic fibroblasts (MEFs) we isolated human fetal liver stromal cells (hFLSCs) from 14 weeks human fetal liver as new human feeder cells. hFLSCs feeders could maintain hES cells for 15 passages (about 100 days). Basic fibroblast growth factor (bFGF) is known to play an important role in promoting self-renewal of human embryonic stem (hES) cells. So, we established transgenic hFLSCs that stably express bFGF by lentiviral vectors. These transgenic human feeder cells--bFGF-hFLSCs maintained the properties of H9 hES cells without supplementing with any exogenous growth factors. H9 hES cells culturing under these conditions maintained all hES cell features after prolonged culture, including the developmental potential to differentiate into representative tissues of all three embryonic germ layers, unlimited and undifferentiated proliferative ability, and maintenance of normal karyotype. Our results demonstrated that bFGF-hFLSCs feeder cells were central to establishing the signaling network among bFGF, insulin-like growth factor 2 (IGF-2), and transforming growth factor β (TGF-β), thereby providing the framework in which hES cells were instructed to self-renew or to differentiate. We also found that the conditioned medium of bFGF-hFLSCs could maintain the H9 hES cells under feeder-free conditions without supplementing with bFGF. Taken together, bFGF-hFLSCs had great potential as feeders for maintaining pluripotent hES cell lines more safely and economically.
authors.affiliation: Stem Cell and Regenerative Medicine Lab, Beijing Institute of Transfusion Medicine, Beijing, China.
authors.forename: Jiafei
authors.initials: J
authors.lastname: Xi
authors.forename: Yunfang
authors.initials: Y
authors.lastname: Wang
authors.forename: Peng
authors.initials: P
authors.lastname: Zhang
authors.forename: Lijuan
authors.initials: L
authors.lastname: He
authors.forename: Xue
authors.initials: X
authors.lastname: Nan
authors.forename: Wen
authors.initials: W
authors.lastname: Yue
authors.forename: Xuetao
authors.initials: X
authors.lastname: Pei
mesh_terms: D002454:Cell Differentiation; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D053595:Embryonic Stem Cells / Q000378:metabolism*; D016222:Fibroblast Growth Factor 2 / Q000096:biosynthesis*; D006801:Humans; D007335:Insulin-Like Growth Factor II / Q000378:metabolism; D007621:Karyotyping; D016086:Lentivirus / Q000235:genetics; D008099:Liver / Q000196:embryology*; D039904:Pluripotent Stem Cells / Q000166:cytology; D017154:Stromal Cells / Q000378:metabolism*; D013724:Teratoma / Q000378:metabolism; D016212:Transforming Growth Factor beta / Q000378:metabolism; D019076:Transgenes
chemical_list: D016212:Transforming Growth Factor beta; D016222:Fibroblast Growth Factor 2; D007335:Insulin-Like Growth Factor II