Culture Of Rat Bone Marrow Derived Neural Tissue Committed Stem Cell And Study On It’s Tansplantation In Ischemia Rat Model

Objective：To investigate the distribution and differentiation of neuraltissue committed stem cells by vein transplantation in middle cerebral arteryocclusion.Methods: Monocyte were collected from bone marrow and culturedin DMEM/F12serum free medium plus EGF、bFGF、2%B27to get Neuraltissue committed stem cells (NTCSCs). Expression of Nestin of NTCSCs，were identified by immunocytochemistry.The differentiation were checked.Modified Zea-Longa suture method were used to make middle cerebralartery occlusion (MCAO) model. The NTCSCs were labeled by DAPI andwere injected through caudal vein of rats; At1,3,5,7days after injection ratswere sacrificed and the brian tissue of transplantation group were preparedas frozen section.The distribution of transplanted NTCSCs was observedunder fluorescent microscope and the differentiation of transplantedNTCSCs were observed under laser confocal microscopy afterimmunofluorescent staining. Results: The cultured NTCSC formed NSC spheres in Nsc culturemedium and expressed Nestin and CXCR4at protein level; RT-PCR showedNTCSCs expressed Nestin、CXCR4、CD31、β-Ⅲ-tubulin、GFAP mRNA;After transplantation the DAPI labelled NTCSCs could be observed in thebrain at3days and a few of cells expressing GFAP could be found at7days.Conclusion: The NTCSCs by intravenous transplantation can enter ratbrain，mainly distributed in ischemia injury，some NTCSCs labelled withDAPI in brain tissue could express GFAP.