Experimental Studies Of Bone Marrow Mesenchymal Stem Cells Transplantation In Intracerebral Hemorrhage

【Objective】To observe the different effects on the recovery of motor and somatosensory function of rats by administrating hBMSCs into rats via the different way of subarachnoid space , tail intravenous and intracerebral injection.【Methods】1. hBMSCs were collected, and after density gradient centrifugation and plastic plate adherent culture, these cells were identified by immuno-fluorescence staining and flow cytometric analysis (FACS) and inducing differentiation to fatcell and neuron-like cells.2. Experimental ICH was induced by intrastriatal administration of unheparinized autoblood from rat. Experimental SD rats were randomly divided into five groups: A, Sham operation group (n=8); B, the negative control (n=10); C, the group directly administrate hBMSCs by stereotaxis injecting into the cerebra (n=10); D, the intravenous hBMSCs transplantation group (n=10); E, subarachnoid-space hBMSCs transplantation group (n=10).3. hBMSC were labled with Hoechst33342 and transplanted into intracerebral hemorrhage rats 4-7 days after the model were setted up.The neurological motor and somatosensory function were assessed by Menzies 5 grade criterion and adhesive-removel somatosensory test at pre-transplanted and post-transplanted . The frozen sections of the rats'cerebra were obtained on the 14th and 28th day after the transplantation respectively. Fluorescence microscope was used to observe the distribution of the marked cells in the brains of the rats.【Results】1.The havestded cells by density gradient centrifugation and plastic plate adherent , 14 days after culture , like elements forming colonies of fibroblastoid cells, they expressed CD106,CD105,CD29, but not CD34,CD45, and induced differentiation to fatcell and neuron-like cells by different inductor.2. hBMSCs were observed to migrate to the area around the lesion site by Fluorescence microscope both in C , D and E groups . A large number of HBMSCs could be seen within 2 weeks after transplantion and few cells could be found at the end of 1 month .3. Functional status was balanced between the HBMSCs-treated groups and the control at baseline, and no significant difference on each behavioral score (Menzies and adhesive-removel patch test) was deteched with P>0.05.4. Menzies 5 grade criterion score showed that the mean score of C and E lower than B 5 days after transplantion , P<0.05 . no significant difference between B and D , C and E . Rats with HBMSCs treatment groups had significant functional recovery at 14 days and 28days compared with rats in the control group with P<0.05. no significant difference among MSCs treatment groups , P>0.05 .5. Adhesive-removel somatosensory test showed that the mean latency period Rats both 5 days and 14 days after transplantion of hBMSCs treatment groups were significant shorter than rats in the control group with P<0.05 , The significant difference persisted until 28 days after transplantion .the mean latency period of Sham operation group was the shortest , P<0.05 .6. No neoplasia was found in the brain of the host 28 days after transplantation.【Conclusions】1.hBMSCs could transcend the blood- brain barrier and migrate to the area around the lesion site , and survive for a certain period .2.hBMSCs transplantation could accelerate the recover of the neurologic impairment.3 . hBMSCs could improve the neurological function of rats with ICH via subarachnoid space liking transplanting directly by stereotaxis .4.Adverse reaction of HBMSCs transplantion was not observed in our data , but required long-term safety evaluation studies .