【摘要】：目的：研究胚胎神經(jīng)干細(xì)胞和聽(tīng)囊細(xì)胞體外分化為毛細(xì)胞的潛能；探討胚胎神經(jīng)干細(xì)胞在內(nèi)耳再生研究領(lǐng)域的價(jià)值。 方法：分離孕9.5-10.5天C5786/L小鼠胚胎的后腦區(qū)域的神經(jīng)管組織和聽(tīng)囊組織,采用無(wú)血清培養(yǎng),進(jìn)行體外培養(yǎng)、擴(kuò)增,并通過(guò)免疫熒光化學(xué)染色(巢蛋白)進(jìn)行鑒定。取傳至第4代神經(jīng)干細(xì)胞和聽(tīng)囊細(xì)胞,接種于蓋玻片上。添加血清培養(yǎng)基,培養(yǎng)15天后行免疫熒光檢測(cè),與RT-PCR檢測(cè)。觀察神經(jīng)干細(xì)胞和聽(tīng)囊細(xì)胞的體外分化特點(diǎn)。 結(jié)果：采用無(wú)血清培養(yǎng)法成功地培養(yǎng)出胚胎神經(jīng)干細(xì)胞球和聽(tīng)囊細(xì)胞球,采用神經(jīng)球分析法收集細(xì)胞球進(jìn)行純化與傳代,免疫熒光檢測(cè)示：來(lái)源于神經(jīng)管和聽(tīng)囊的細(xì)胞球巢蛋白(nestin)表達(dá)陽(yáng)性,證實(shí)其都具有神經(jīng)干細(xì)胞的特點(diǎn)。添加含血清培養(yǎng)基培養(yǎng)15天后,免疫熒光檢測(cè)證實(shí)所獲得的神經(jīng)干細(xì)胞和聽(tīng)囊細(xì)胞均可以在體外分化為表達(dá)毛細(xì)胞標(biāo)記物MyosinV Ⅶa的細(xì)胞。 結(jié)論：胚胎神經(jīng)干細(xì)胞和聽(tīng)囊細(xì)胞能夠在體外培養(yǎng)的條件下分化為毛細(xì)胞樣細(xì)胞：胚胎神經(jīng)干細(xì)胞可以用于研究?jī)?nèi)耳毛細(xì)胞分化過(guò)程中的重要基因表達(dá)與調(diào)控因子、揭示聾病發(fā)生的機(jī)制及耳蝸的細(xì)胞移植治療的可行性。
[Abstract]:Aim: to study the potential of embryonic neural stem cells and auditory follicle cells to differentiate into hair cells in vitro and to explore the value of embryonic neural stem cells in the field of inner ear regeneration. Methods: the neural tube tissues and auditory bursa tissues in the posterior brain of 9.5-10.5 days C5786 / L mouse embryos were isolated, cultured in vitro and amplified by serum-free culture, and identified by immunofluorescence staining (nestin). Neural stem cells and auditory sac cells were transferred to the fourth passage and inoculated on the cover glass. After 15 days of culture, serum was added to the medium and immunofluorescence assay was performed with RT-PCR. To observe the differentiation characteristics of neural stem cells and auditory sac cells in vitro. Results: embryonic neural stem cell ball and auditory sac cell ball were successfully cultured by serum-free culture. The expression of nestin (nestin) was positive in neurons from nerve tube and auditory sac, which confirmed that they had the characteristics of neural stem cells. After cultured in serum-containing medium for 15 days, the obtained neural stem cells and auditory sac cells were confirmed to be able to differentiate into cells expressing hair cell marker MyosinV 鈪，

本文編號(hào)：2199333