【摘要】： 目的對(duì)大鼠陰莖海綿體細(xì)胞進(jìn)行初步分離培養(yǎng),檢測(cè)干細(xì)胞標(biāo)志物干細(xì)胞抗原-1(Sca-1)、Oct4和肌源性標(biāo)志物結(jié)蛋白(Desmin)的表達(dá),探索陰莖海綿體中是否存在內(nèi)源性干細(xì)胞及肌源性干細(xì)胞。 方法取2月齡SD大鼠,重量200～250g,乙醚吸入麻醉后,無(wú)菌條件下取陰莖海綿體組織,用Ⅰ型膠原酶、胰蛋白酶消化分離組織,待細(xì)胞基本以單細(xì)胞形態(tài)存在時(shí),過(guò)濾、離心收集細(xì)胞沉淀,用含20!胎牛血清的DMEM重懸培養(yǎng)。采用改良的Preplate差速貼壁技術(shù)對(duì)細(xì)胞進(jìn)行初步分離培養(yǎng),所獲PP1-PP6貼壁細(xì)胞,應(yīng)用流式細(xì)胞儀、免疫熒光細(xì)胞化學(xué)法、western blot檢測(cè)觀察分離細(xì)胞中Sca-1、Oct4及Desmin的表達(dá),同時(shí)行Sca-1/Oct4、Sca-1/Desmin、Oct4/Desmin免疫熒光雙標(biāo)記。 結(jié)果分離培養(yǎng)所獲PP1和PP2細(xì)胞貼壁快,主要是長(zhǎng)梭形的成纖維樣細(xì)胞,PP3-PP5細(xì)胞貼壁能力逐漸減慢,出現(xiàn)短梭形細(xì)胞,有的呈多角形,以血管內(nèi)皮細(xì)胞、平滑肌細(xì)胞為主,PP6細(xì)胞中有體積較小,成圓形的漂浮細(xì)胞,這種細(xì)胞貼壁很慢,2-3天后開(kāi)始貼壁成圓形或梭形。流式細(xì)胞儀檢測(cè):PP3細(xì)胞中Sca-1、Oct4陽(yáng)性表達(dá)量分別為0.3!±0.2!、0.2!±0.3!,與同型對(duì)照組相比均無(wú)統(tǒng)計(jì)學(xué)差異(p㧐0.05),Desmin陽(yáng)性表達(dá)量為15.3!±1.5!,與同型對(duì)照組相比有統(tǒng)計(jì)學(xué)差異(p㩳0.05);PP6細(xì)胞中Sca-1、Oct4、Desmin陽(yáng)性表達(dá)量分別為5.7!±0.7!、2.6!±0.3!、41.1!±1.6!,與PP3相比均有統(tǒng)計(jì)學(xué)差異(p㩳0.05)。免疫熒光細(xì)胞化學(xué)顯示在PP6中有少量細(xì)胞分別表達(dá)Sca-1和Oct4,均散在分布,Sca-1以胞漿表達(dá)為主,Oct4則表達(dá)于胞核;表達(dá)Desmin細(xì)胞聚集,數(shù)量較多,主要表達(dá)于胞漿;亦發(fā)現(xiàn)有極少量雙陽(yáng)性標(biāo)記細(xì)胞(Sca-1/Oct4、Sca-1/Desmin和Oct4/Desmin),Sca-1/Oct4表達(dá)于同一細(xì)胞的胞漿及胞核,Sca-1/Desmin表達(dá)于同一細(xì)胞的胞漿,Oct4/Desmin則表達(dá)于同一細(xì)胞的胞核及胞漿。Western blot顯示PP1-PP5細(xì)胞中未檢測(cè)到Sca-1及Oct4蛋白明顯表達(dá),但在PP6細(xì)胞中則檢測(cè)到Sca-1及Oct4蛋白的表達(dá),Desmin蛋白在PP1-PP6細(xì)胞中表達(dá)逐漸增強(qiáng)。 結(jié)論在大鼠陰莖海綿體中發(fā)現(xiàn)具有干細(xì)胞及肌源性干細(xì)胞表型的細(xì)胞,提示陰莖海綿體中存在內(nèi)源性干細(xì)胞及肌源性干細(xì)胞。
[Abstract]:Objective to investigate the expression of stem cell antigen 1 (Sca-1), Oct4 and desmin (Desmin) in rat penis cavernous cells. To explore the existence of endogenous stem cells and myogenic stem cells in the cavernous body of penis. Methods two month old SD rats, weighing 200,250g, were taken from the corpus cavernosum of penis under aseptic condition after inhaled anesthesia with ether. The isolated tissues were digested by type 鈪，

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