本文選題：人臍帶間充質干細胞 + 神經(jīng)干細胞�。� 參考：《聽力學及言語疾病雜志》2017年03期

【摘要】：目的探討并建立一種體外定向誘導人臍帶間充質干細胞(human umbilical cord mesenchymal stem cells,hUC-MSCs)向內耳毛細胞樣細胞分化的方法。方法采用組織貼壁法培養(yǎng)獲得hUC-MSCs,流式細胞儀鑒定其表面標記物;神經(jīng)干細胞誘導階段分為對照組(DFNB基礎誘導培養(yǎng)基)、實驗1組(DFNB基礎誘導培養(yǎng)基+20ng/ml EGF+20ng/ml bFGF)、實驗2組(DFNB基礎誘導培養(yǎng)基+20ng/ml EGF+20ng/ml bFGF+瓊脂糖包被),培養(yǎng)3~5天后采用免疫熒光法檢測各組標記蛋白Nestin的表達;內耳毛細胞樣細胞誘導分化階段,分為對照組(DFNB基礎誘導培養(yǎng)基+明膠包被)、誘導組[DFNB基礎誘導培養(yǎng)基+20ng/ml EGF+1μM全反式維甲酸/全反式維生素A酸(all trans retinoic aid,ATRA)],培養(yǎng)4周后采用qRT-PCR和免疫熒光法檢測各組毛細胞標記物Math1、MyosinⅦa、Brn3c的表達。結果在神經(jīng)干細胞誘導階段,對照組、實驗1組均無神經(jīng)球結構,實驗2組中hUC-MSCs分化3d后細胞有明顯神經(jīng)球樣結構,且Nestin陽性細胞數(shù)更高(P0.05);而向內耳毛細胞樣誘導分化4周后,誘導組毛細胞標記物Math1、MyosinⅦa、Brn3c的mRNA和陽性細胞數(shù)的表達水平明顯升高(P0.05)。結論體外誘導人臍帶間充質干細胞先向神經(jīng)干細胞方向分化再向內耳毛細胞分化方向誘導,可有效提高毛細胞標記物MyosinⅦa、Brn3c、Math1的表達水平,促進hUC-MSCs向類毛細胞樣細胞分化,可能為將來感音神經(jīng)性聾的內耳移植治療提供更為適合的種子細胞。
[Abstract]:Objective to explore and establish a method for inducing the differentiation of human umbilical cord mesenchymal stem cells from human umbilical cord mesenchymal stem cells into inner ear hair cell-like cells in vitro. Methods hUC-MSCs were obtained by tissue adherence method and their surface markers were identified by flow cytometry. The neural stem cells were divided into two groups: control group (control group), DFNB basic induction medium (20ng/ml EGF 20ng/ml bFGFN), experimental group 1 (20ng/ml EGF 20ng/ml bFGFN), and experimental group 2 (20ng/ml EGF 20ng/ml bFGF agarose coated medium), which were cultured for 3 days and then used for 5 days. The expression of labeled protein Nestin in each group was detected by immunofluorescence assay. The differentiation of hair cell-like cells in inner ear was induced. The cells were divided into two groups: the control group (20ng/ml EGF 1 渭 M 20ng/ml EGF 1 渭 M) and the control group (20ng/ml EGF 1 渭 M, 20ng/ml EGF 1 渭 M). After 4 weeks of culture, qRT-PCR and immunofluorescence were used to detect each group. The expression of the hair cell marker Math1, Myosin 鈪，

本文編號：1937048