Migration and Neuron Differentiation of Neural Stem Cells When Transplanted to the Lateral Ventricle of Neonatal Rat / 实用儿科临床杂志

ABSTRACT

Objective To explore the migration and differentiation of the human neural stem cells (hNSC) after being transplanted to the neonatal rat lateral ventricle,to provide some data on therapy for neonatal cerebropathy by using of neural stem cells.Methods 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).The part of the hNSC in-vitro cultured for 14 d was digested to suspensions of cell.Cultured for 14 d, the hNSC in-vitro and the suspension were transplanted into the lateral ventricles of the neonatal rat brains.The rats were 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).Results In vitro culture,the typical NSC spheres were obtained from the forebrain of the human fetus.The suspensions of cells were obtained from the neurosphere.In neurosphere group, the results of anti-hNuc detecting tracing at 72 h post-injection showed that the grafts had migrated into the cortex grand layers of olfactory bulbus,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.In suspensions group,the results of anti-hNuc detecting tracing at 24 h post-injection 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.Conclusions Whole neurospheres migrated intensely and differentiated into neurons and gliocytes.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.