In our present study, we aimed at determining whether the hES cells can be maintained pluripotent using bFGF-hFLSCs-CM rather than xenogenic MEF-CM. Similar to feeder-free culture in MEF-CM and mTeSR1, H9 hES cells can be equally maintained stable and pluripotent for over 15 passages. H9 hES cells maintained in bFGF-hFLSCs-CM without supplementing with bFGF retained typical morphology, expression of surface markers and transcription factors associated with pluripotency The conditioned medium of bFGF-hFLSCs could be used to develop a new hES cell feeder-free culture condition. In conclusion, we isolated a new human feeder cell for maintenance of H9 hES cells. And then, we established transgenic cell lines �� bFGF-hFLSCs that stably express bFGF, which could be used as feeder cells culturing H9 hES cells without supplementing with exogenous growth factors. More importantly of the bFGF-hFLSCs-CM could maintain H9 hES cell pluripotency in feeder-free culture conditions. bFGF-hFLSCs had great potential as feeders for maintaining hES cells more safely and economically. Mesenchymal stem cells are multipotent cells present in the stroma of most organs. Their isolation relies on adherence to cell culture plastics. In culture, human MSCs are capable of extensive proliferation without giving rise to chromosomal aberrations. Large numbers of hMSCs can thus be obtained from small samples of easily accessible tissues like fat and bone marrow. MSCs are identified primarily on the basis of their ability to differentiate under specific culture Z-VAD-FMK conditions into various mesodermal cell types including osteoblasts, chondrocytes and adipocytes. Since markers exclusively displayed on the surface of MSCs have not been identified yet, these cells are phenotypically characterized by a Masitinib combination of several non-hematopoietic cell surface markers. Typically, cultured hMSCs express on their surface the major histocompatibility complex class I human leukocyte antigens A, B, C and HLA-G, but not HLA-E 3, 4 and our unpublished observations. Furthermore, hMSCs do not display MHC class II proteins on their plasma membrane. MHC class II molecules are present, however, in intracellular reservoirs and can be recruited to the cell surface through exposure to interferon gamma. Recently, cultured MSCs were shown to have pleiotropic immunomodulatory properties in vitro including suppression of T cell proliferation in response to alloantigens or mitogens, inhibition of B cell proliferation and antibody production and inhibition of dendritic cell maturation.