The Immunophenotypes Of Inner Ear Hair Cell From Guine Pig Central Nervous System In Vitro And Explantation In Vivo

Division, Culture and Identification of Neural Stem Cells form the Guinea PigObjective Division,culture and identification of neural stem cells form the guinea pig to creat bases for the experimental study on treating sensorinerual deafness by transplantation of neural stem cells.Method The brain get from the newborn guinea pig brain tissues were disassociated and trypsinizated in the D-Hank's solution. Cells were cultuled in the media composed of 20% B27,20ng/ml EGF,20ng/ml bFGF and DMEM/F12. Neural stem cells were identified by Nestin immunohistochemistry. Then the cells were serial subcultivated.Results Clone like cluster of neural stem cells were obtained from the whole brain. Most of the cells expressed nestin epitopes,but not Brn-3c or myosinⅦaConclusions the cells from brains of fetal rat have the potential of proliferation. They can be used as donor cells to experimental research on treating neurosensory deafness with transplantation of neural stem cells. The Immunophenotypes of Inner Ear Hair Cell from Guinea Pig Central Nervous System In VitroObjective To exame whether hair cell immunophenotypes could be derived from the Central Nervous System(CNS) in vitro.Methods The neural stem cells were cultuled in the media composed of 20% B27,20ng/ml EGF,20ng/ml bFGF,TGF-α,IGF-Ⅰ,10% FBS and DMEM/F12 for four weeks. Then we use the immunohistochemistry to identified the cells with the antibody of myosinⅦa,pan-cytokeratin。Results The cells expressed immunoreactivity for either myosinⅦa or pan-cytokeratin.Conclusions The neural stem cells posses the potential to differentiate into hair cell phenotypes in vitro. That provide the means to explore therapeutic methods of hearing dysfunction by cellular transplant with the neural stem cells. Preliminary Study of Neural Stem Cell Transplantation into the Guinea Pig CochleaObjective To establish the method of neural stem cell (NSC) transplantation into guinea pig cochlea and analyze the feasibility of NSC transplantation.Methods NSCs were isolated from the brain of the newborn guinea pig in vitro. NSCs labeled with Hoechst33342 were transplanted into the guinea pig cochlea via a small hole drilled on the bony wall of tympanic scala in the basal turn of the cochlea. The cell suspension was inoculated into the perilymphatic space, and the culture fluid without cell suspension was injected as a negative control. After 7-10 days the animals were killed, and the cochlea were cut into a frozen section. The transplanted cells were examined with fluorescent microscope. Some of frozen sections were observed by HE staining.Result The transplanted cells were survived along the bone of the cochlea.Conclusion NSC can survive in the inner ear transplantation, which provides a basis for study of NSC differentiation and expression in inner ear.