In-vitro experimental study on Atoh1 gene inducing differentiation of rat dermal papilla cells to inner ear hair cells

Abstract

Objective To investigate the expression and biological effect of Atoh1 in dermal papilla cells (DPCs) in rats,so as to provide a new treatment for sensorineural deafness.Methods Atoh1 was transfected into rat DPCs using lentivirus infection method and co-cuhured with the organ of Corti in vitro.Expression of Atoh1 mRNA in DPCs was detected with RT-PCR; lentivirus transduction efficiency and morphological changes of DPCs were observed using microscope ; expression of DPCs specific antibodies was measured with immunohistochemical staining.Results Transfection efficiency was (87.3 +3.1) ％ at 48 hours posttransduction and typical hair-cell like cells increased under the microscope.Immunohistochemical analysis showed cells with positively expressed nestin protein,neuron special endolase (NSE),and glial fibrillary acidic protein (GFAP) reached (43.5 ± 2.5) ％,(31.3 ± 2.4) ％,and (10.3 ± 2.9) ％ respectively.Positive rate of hair cell-specific protein (myosin protein Ⅶa,MYOⅦa) was (13.2 ± 1.3) ％.Conclusion Atoh1 gene co-cultured with the organ of Corti in vitro can promote the differentiation of DPCs to cochlear hair cells,which provides a theoretical basis for treating sensorineural deafness by dermis-derived stem cell transplantation. Key words: Hair cells, auditory, inner; Regeneration; Gene therapy