中华创伤杂志
中華創傷雜誌
중화창상잡지
Chinese Journal of Traumatology
2014年
8期
822-826
,共5页
朱珠%郑国玺%祝康%罗娟%张璐瑶%夏翠%韦俊荣%许珉
硃珠%鄭國璽%祝康%囉娟%張璐瑤%夏翠%韋俊榮%許珉
주주%정국새%축강%라연%장로요%하취%위준영%허민
毛细胞,听觉,内%再生%基因治疗
毛細胞,聽覺,內%再生%基因治療
모세포,은각,내%재생%기인치료
Hair cells,auditory,inner%Regeneration%Gene therapy
目的 探讨Atoh1基因在大鼠真皮毛乳头细胞(dermal papilla cells,DPCs)中的表达及生物学作用,为治疗感音神经性耳聋建立一种新的方法. 方法 利用慢病毒介导Atoh1基因感染大鼠DPCs,并联合体外耳蜗Corti器共培养.RT-PCR检测Atoh1 mRNA的表达,显微镜下观察慢病毒感染效率及DPCs形态学变化,免疫组化法检测DPCs细胞特异性抗体的表达. 结果 Atoh1感染DPCs 48 h后的转染效率为(87.3±3.1)％,显微镜下可见典型类毛细胞形态样细胞增多;免疫组化显示转染后细胞巢蛋白、神经元特异性烯醇化酶(neuron special endolase,NSE)、胶质纤维酸性蛋白(glial fibrillary acidic protein,GFAP)阳性细胞率分别为(43.5±2.5)％、(31.3±2.4)％和(10.3±2.9)％;毛细胞特异性肌球蛋白(myosin proteinⅦa,MYOⅦa)表达阳性细胞率为(13.2±1.3)％. 结论 Atoh1基因联合体外耳蜗Corti器共培养可促进DPCs向耳蜗毛细胞方向分化,为真皮源性干细胞引入内耳治疗感音神经性耳聋提供实验依据.
目的 探討Atoh1基因在大鼠真皮毛乳頭細胞(dermal papilla cells,DPCs)中的錶達及生物學作用,為治療感音神經性耳聾建立一種新的方法. 方法 利用慢病毒介導Atoh1基因感染大鼠DPCs,併聯閤體外耳蝸Corti器共培養.RT-PCR檢測Atoh1 mRNA的錶達,顯微鏡下觀察慢病毒感染效率及DPCs形態學變化,免疫組化法檢測DPCs細胞特異性抗體的錶達. 結果 Atoh1感染DPCs 48 h後的轉染效率為(87.3±3.1)％,顯微鏡下可見典型類毛細胞形態樣細胞增多;免疫組化顯示轉染後細胞巢蛋白、神經元特異性烯醇化酶(neuron special endolase,NSE)、膠質纖維痠性蛋白(glial fibrillary acidic protein,GFAP)暘性細胞率分彆為(43.5±2.5)％、(31.3±2.4)％和(10.3±2.9)％;毛細胞特異性肌毬蛋白(myosin proteinⅦa,MYOⅦa)錶達暘性細胞率為(13.2±1.3)％. 結論 Atoh1基因聯閤體外耳蝸Corti器共培養可促進DPCs嚮耳蝸毛細胞方嚮分化,為真皮源性榦細胞引入內耳治療感音神經性耳聾提供實驗依據.
목적 탐토Atoh1기인재대서진피모유두세포(dermal papilla cells,DPCs)중적표체급생물학작용,위치료감음신경성이롱건립일충신적방법. 방법 이용만병독개도Atoh1기인감염대서DPCs,병연합체외이와Corti기공배양.RT-PCR검측Atoh1 mRNA적표체,현미경하관찰만병독감염효솔급DPCs형태학변화,면역조화법검측DPCs세포특이성항체적표체. 결과 Atoh1감염DPCs 48 h후적전염효솔위(87.3±3.1)％,현미경하가견전형류모세포형태양세포증다;면역조화현시전염후세포소단백、신경원특이성희순화매(neuron special endolase,NSE)、효질섬유산성단백(glial fibrillary acidic protein,GFAP)양성세포솔분별위(43.5±2.5)％、(31.3±2.4)％화(10.3±2.9)％;모세포특이성기구단백(myosin proteinⅦa,MYOⅦa)표체양성세포솔위(13.2±1.3)％. 결론 Atoh1기인연합체외이와Corti기공배양가촉진DPCs향이와모세포방향분화,위진피원성간세포인입내이치료감음신경성이롱제공실험의거.
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.
