Distribution and differentiation of transplanted neural tissue committed stem cells to the brain after ischemic stroke / 解放军医学杂志

ABSTRACT

Objective To investigate the distribution and differentiation of neural tissue committed stem cells (NTCSCs) by venous transplantation to the brain after middle cerebral artery occlusion in rats. Methods Monocytes were collected from bone marrow and cultured in DMEM/F12 serum free medium plus EGF, bFGF and 2% B27 to obtain NTCSCs. Expressions of nestin, CXCR4, CD31, β -HI-tubulin and GFAP mRNA in NTCSCs were detected by RT-PCR. Modified Zea-Longa suture method was used to prepare middle cerebral artery occlusion (MCAO) model. The NTCSCs were labeled with DAPI and were injected through the caudal vein of the rats. At 1,3,5,7 days after injection, the rats were sacrificed and the brain tissues were prepared with frozen section in transplantation group. The distribution and differentiation of transplanted NTCSCs were observed under fluorescent microscope and laser confocal microscope respectively after immunofluorescent staining. Results The cultured NTCSC formed neural stem cell spheres in neural stem cell culture medium, and they expressed Nestin and SDF-1 receptor CXCR4 at protein level. RT-PCR showed that NTCSCs expressed Nestin, CXCR4, CD31, β - III-tubulin and GFAP mRNA. DAPI-labelled NTCSCs could be observed in the brain 3 days after transplantation and a few GFAP expressing cells could be found on the 7th day. Conclusion The NTCSCs by intravenous transplantation can enter rat brain, and they distribute mainly in ischemic tissues. Some NTCSCs labelled with DAPI in the brain tissue could express GFAP.