Homing Study Of Human Embryonic Stem Cells Derived Mesenchymal Stem Cells And Surface Marker Study Of Embryonic Stem Cells

Embryonic stem cells (ES) possess unlimited proliferation potency in vitro, committed differentiated ES could be an excellent resource for cell transplantation. Studies of homing behaviors of committed differentiated ES in pathological models are important for therapeutic application of ES. In the present study, human ES were first induced to MSC by treating with inhibitor of ROCK and further cultured in medium supplemented with serum. Then hES-MSC were labeled with biotin for in vivo tracking.36h after hES-MSC were xenogenic transplanted into ICR mouse, flow cytometry and immunohistochemistry was carried out and results showed that hES-MSC would more likely home to bone marrow, spleens and lungs but not livers in normal ICR mouse, and a large number of hES-MSC could be detected in livers of acute liver injured ICR mouse. Passage numbers would significantly affect early homing of hES-MSC in normal ICR mouse, but the ability of hES-MSC homing to injured liver was almost the same despite of the difference of passage number. Further experiments showed that CXCR4 played an important role in hES-MSC early homing to injured liver, and milieu of injured liver could promote expressions of CXCR4 in hES-MSC. Hepatic differentiation of hES-MSC homed to injured liver was also observed, which AFP was highly expressed in these hES-MSC. We also found that TNF-αcould promote expressions of CXCR4 in hES-MSC, and hES-MSC pre-incubated with TNF-αcould home to injured liver more effectively. Hence, we may conclude that hES-MSC could home to injured liver and start hepatic differentiation in ICR mouse at early stage of cell transplantation, and TNF-αcould be a potent candidate for enhancing hES-MSC early homing to injured liver.Surface markers of mouse embryonic stem cells (mES), correlated with signal transduction and intercellular signal transmission, are important for self renew and differentiation of mES. In the present study, rabbit monoclonal antibodies against native form of mouse ES surface markers were generated. Then the target antigens were purified by improved immunoprecipitation, and further identified by LC-LTQ mass spectrum and confirmed by western blot using commercial-available antibodies. At last a mAb against native form of Glut3 on outer membrane of mES was obtained, and further experiments showed that blockage of Glut3 by using mAb obtained in our experiments could inhibit proliferation of mES, and indicated that Glut3 are important for self renew of mES.