Migration And Differentiation Of Human Embryo Neural Stem Cells When Transplanted To The Lateral Ventricle Of Neonatal Rat

Objective To explore the migration and differentiation of the human neural stem cells(hNSC) after being transplanted to the neonatal rat lateral ventricle.To compare the different destiny of Whole neurospheres and cells from suspension after transplanting to the brain.To provide some data on therapy for neonatal cerebropathy by using of neural stem cells.Material and Method1.Cell culture N2 medium containing EGF+FGF2+LIF was used to culture the NSC spheres from the forebrain tissues of aborted human fetus.The hNSC was identified by detecting the NSC marker nestin antigen and showing the potency to differentiate into neural cells(including astrocytes,oligodendrocytes and neurons) by using indirect immuno - fluorescence assay(IFA).2.Animal and Subgroups Thirty-six Spraquc-Dawley(SD) rats were randomly divide into experiment group(NSC spheres group,cell suspension group) and control group(F12 culture solution group).3.The ratswere respectively killed at 24,48 and 72 h respectively post - transplant, the whole brain was sectioned,and the special immuno - response detection was performed by using anti - human nuclei(anti - hNuc) and anti - human neurofilament (anti-hNF).Results1.In vitro culture,the typical NSC sphereswere obtained from the forebrain of the human fetus.Immunofluorescence sign that the NSC has the potential of differentiating into the neuron and glial cell. 2.Two different fates between NSC spheres and cell suspension in the host rat brains. NSC spheres group:The results of anti - hNuc detecting tracing at 72 h postinjection showed that the grafts had migrated into the cortex grand layers of olfactory bul-bus,medial precentral area of lobus frontalis,hippocampal,and lobus occipitalis. The label - positive cells lined along the Cerebellar Purkinje cell layers and appeared in most parts of mesencephalons.The immuno - respons results of anti - hNF showed that the positive cells scattered in the grand layer of cortex,the connection among positive cells was watched.cell suspension group:the results of anti - hNuc detecting tracing at 24 h postinjection show a great quantity of positive cells in the ventricles and injection track. At 72 h,a small quantity of positive cells remained in the ventricle and nearby brain tissue.Conclusion1.Whole neurospheresmigrated intensely and differentiated into neurons and gliocytes.2.At the same time,transplants of cells from suspension transplants showed limited or no migration because of internal environment of the brain and construction of neurospheres.