【摘要】：目的為應(yīng)用神經(jīng)生長因子(NGF)修飾的骨髓基質(zhì)干細(xì)胞(BMSC)進(jìn)行耳蝸移植治療老年性耳聾的實(shí)驗研究提供支持。方法采用脂質(zhì)體LipofectamineTM2000方法進(jìn)行骨髓基質(zhì)干細(xì)胞轉(zhuǎn)染,共分為4組:A組:脂質(zhì)體包裹的重組質(zhì)粒轉(zhuǎn)染組;B組:脂質(zhì)體轉(zhuǎn)染組;C組:脂質(zhì)體包裹的空質(zhì)粒轉(zhuǎn)染組;D組:用含10%胎牛血清的DMEM F12培養(yǎng)液培養(yǎng)。免疫組化檢測各組轉(zhuǎn)染后神經(jīng)生長因子和神經(jīng)生長因子受體p75的表達(dá)。結(jié)果各轉(zhuǎn)染組細(xì)胞均有NGF、受體p75表達(dá),且均以胞漿表達(dá)為主;圖像分析顯示,重組質(zhì)粒組NGF表達(dá)的平均光密度值明顯高于其余3組,重組質(zhì)粒組受體p75表達(dá)的平均光密度值明顯低于其余3組。結(jié)論采用脂質(zhì)體LipofectamineTM2000轉(zhuǎn)染方法能成功將含有NGF的重組質(zhì)粒轉(zhuǎn)染至BMSC,為我們進(jìn)一步應(yīng)用NGF修飾的BMSC進(jìn)行耳蝸移植治療老年性耳聾的實(shí)驗研究提供理論基礎(chǔ)。
[Abstract]:Objective to provide support for cochlear transplantation with bone marrow stromal stem cells (BMSC) modified by nerve growth factor (NGF) in the treatment of senile deafness. Methods Bone marrow stromal cells (BMSCs) were transfected by liposome LipofectamineTM2000 method and divided into four groups: group A: liposome-coated recombinant plasmid transfection group; group B: liposome-transfected group; group C: liposome-coated empty plasmid transfection group; Group D: cultured in DMEM F 12 medium containing 10% fetal bovine serum. The expression of nerve growth factor (NGF) and nerve growth factor receptor p75 (NGF receptor p75) were detected by immunohistochemistry. Results the expression of NGF, receptor p75 was found in all the transfected cells, and the expression of p75 was mainly in cytoplasm. Image analysis showed that the average optical density of NGF expression in the recombinant plasmid group was significantly higher than that in the other three groups, and the average optical density of the receptor p75 expression in the recombinant plasmid group was significantly lower than that in the other three groups. Conclusion the recombinant plasmid containing NGF can be successfully transfected into BMSC, by lipofectin LipofectamineTM2000 transfection method, which provides a theoretical basis for the further experimental study of cochlear transplantation with NGF modified BMSC in the treatment of senile deafness.
【作者單位】： 桂林醫(yī)學(xué)院附屬醫(yī)院耳鼻咽喉-頭頸外科;
【基金】：國家自然科學(xué)基金資助項目(編號:81060083) 廣西科學(xué)研究與技術(shù)開發(fā)計劃項目(編號:桂科攻10124001B-56)
【分類號】：R764.436

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