Bone Marrow Mesenchymal Stem Cells Induced To Differentiate Convalescent Therapeutic Effect On Cerebral Experimental Study

Part 1 Study on the effect of ganglioside-1 on differentiation of bone marrowmesenchymal stem cells into neural cellsObjectives: This study aims to explore the effect of ganglioside-1 on thedifferentiatiation of bone marrow mesenchymal stem cells (MSCs) into neural cells.Methods: MSCs were isolated on the basis of its ability to adhere to the culture plate,passaged three times, and finally added GM1 to induce its differentiation. Themorphologic changes of MSCs were observed under phase-contrast microscopy, thenneuron specific enolase (NSE) and glial fibrillary acidic protein (GFAP) expressionwere determined by immunocytochemistry way. NGF mRNA and BDNF mRNAexpression were detected with RT-PCR and then compared with those of FBS controlgroup and DMEM control group.Results: MSCs of GM1 induction group appeared round-shaped, with some dendriteand axon interlaced. (29.47%±3.26) % MSCs showed positive staining to NSE, while(2.32±0.18) % MSCs were positive to GFAP. (6.97±0.56) % and (10.6±0.75) %MSCs were positive to NSE in FBS control group and blank control grouprespectively, while (1.41±0.35) % and (1.21±0.35) % MSCs were positive to GFAP inFBS group and blank control group, respectively. NGF mRNA and BDNF mRNAlevels were significant higher than those of FBS group (P<0.05) and DMEM controlgroup (P<0.01).Conclusions: GM1 can induce MSCs'differentiation into neural cells in vitro. Part 2 Comparison of effectiveness of intravenous or intraperitonealadministration of bone marrow stromal cells on late stage of stroke ratsObjectives: This study was aimed to observe the therapeutic effect of adult bonemarrow stromal cells (MSCs) on late stage of cerebral injury of ischemia, and tocompare the effectiveness between intravenous and intraperitoneal adminstrations ofMSCs so to explore the better way of for the clinical therapy to ischemic stroke, toinvestigate the protective mechanism of MSCs to cerebral ischemic rats afterintraperitoneal administration by comparison with intravenous administration andselect a better administrating path in rats following cerebral ischemia.Methods: Bone marrow cells were isolated, proliferated, purified and passaged fromthe long bone of 180~200g SD rats. MSCs were labeled with BrdU (10mg/L) 24hours before administration. MCAO model was established with modified Zea Longasuture emboli method. Forty-five rats with MCAO were divided into 3 groups atrandom with 15 rats in each group. BrdU labeled-MSCs were injected into MCAOrats through peritoneal and venous paths 3 weeks after model operated, then①Tocompare NSS at different time points (1 day before MCAO operation, at 1 and 21days after MCAO, at 3, 7, 14 and 21 days after MSCs administration).②The ratswere sacrificed at 7, 14 or 21 days after treatment. Their brain tissues were polymethanal-fixed, paraffin-embeded, and paraffin sections.③To evaluate the status ofMSCs, BrdU+MAP-2 and BrdU+GFAP cells from MSCs were analyzed by the meansof immunohistochemistry staining with BrdU, MAP-2 and GFAP as migration ordifferentiation markers.④To evaluate angiogenesis by deteceting the level ofangiogenin expression.⑤To observe MSCs survivorship, migration anddifferentiation in brain tissue after intravenous or intraperitoneal administration.Results: The peak time of NSS in each group is at 1d after administration of MSCs orplacebo. There were no difference among each group (P>0.05). The NSS of intravenous administration groups were lower than that of control groups at 21d afteradministration (P<0.05), while no difference was found between the intraperitonealadministration groups and control groups. Immunohistochemical studies suggestedthat transplanted cells survived and migrated to the ipsilateral cerebral cortex bothafter intraperitoneal and intravenous injection. Some were immunopositive forneuronal marker microtubule associated protein (MAP-2), astrocytic maker glialfibrillary acidic protein (GFAP) or microvessel marker Angiogenin.ConclusionsConclusions: MSCs can migrate into brain tissue to induce neuronagenesis andangiogenesiss either by intraperitoneal or intravenous administration, even at the latestage of ischemic cerebral injuries. The effectiveness of intravenous administration isa better way.