本文選題：牙周膜干細(xì)胞　切入點(diǎn)：D培養(yǎng)　出處：《牙體牙髓牙周病學(xué)雜志》2014年05期 　論文類型：期刊論文

【摘要】：目的:研究懸滴法3D培養(yǎng)和2D貼壁培養(yǎng)的牙周膜干細(xì)胞(PDLSCs)生物學(xué)特性的差異。方法:改良酶消化組織塊法分離培養(yǎng)PDLSCs,分別行2D貼壁培養(yǎng)及3D培養(yǎng)(3D培養(yǎng)按不同細(xì)胞接種數(shù)分為2.5×104,5×104及1×1053組),培養(yǎng)96 h后分別用Live/Dead染色法分析細(xì)胞活性;流式細(xì)胞術(shù)檢測(cè)干細(xì)胞表面標(biāo)記物;然后對(duì)2D、3D培養(yǎng)的PDLSCs進(jìn)行礦化誘導(dǎo),茜素紅染色及熒光定量PCR檢測(cè)礦化結(jié)節(jié)形成情況以及礦化相關(guān)基因ALP和OPN mRNA的表達(dá)水平。結(jié)果:3D培養(yǎng)的PDLSCs可形成致密的細(xì)胞聚合物,2.5×104組PDLSCs細(xì)胞聚合物中心未見(jiàn)壞死現(xiàn)象,5×104組和1×105組則有較為明顯壞死現(xiàn)象;3D和2D培養(yǎng)的PDLSCs間充質(zhì)干細(xì)胞表面標(biāo)記物的陽(yáng)性表達(dá)率均無(wú)顯著差異(P0.05);茜素紅染色和熒光實(shí)時(shí)定量PCR結(jié)果顯示,3D培養(yǎng)的PDLSCs形成礦化結(jié)節(jié)的數(shù)量及其礦化相關(guān)基因ALP OPN mRNA的表達(dá)水平均顯著高于2D培養(yǎng)的細(xì)胞(P0.05)。結(jié)論:3D懸滴培養(yǎng)法是一種理想的PDLSCs培養(yǎng)方法。
[Abstract]:Objective: to study the biological characteristics of periodontal ligament stem cells (PDLSCs) in 3D and 2D adherent culture. Methods: PDLSCs were isolated and cultured by modified enzyme digestion tissue mass method, and were cultured by 2D adherent culture and 3D culture respectively. The cell number was divided into 2.5 脳 10 4 5 脳 10 4 and 1 脳 10 53 groups. After 96 h culture, the cell activity was analyzed by Live/Dead staining. Flow cytometry was used to detect the surface markers of stem cells, and then the PDLSCs cultured in 3D culture was mineralized. Alizarin red staining and fluorescence quantitative PCR were used to detect the formation of mineralized nodules and the expression levels of mineralization-related genes ALP and OPN mRNA. Results the polymer centers of PDLSCs cells in 2.5 脳 10 ~ 4 group of PDLSCs cells were not found to be damaged in the PDLSCs cultured with PDLSCs in 1: 3 D culture. There was no significant difference in the positive expression rate of surface markers of PDLSCs mesenchymal stem cells cultured in 3D and 2D. The results of alizarin red staining and real-time quantitative PCR showed that there was no significant difference between 3D and 2D cultured PDLSCs mesenchymal stem cells. The results of alizarin red staining and fluorescence real-time quantitative PCR showed that there was no significant difference in the expression of surface markers between 3D and 2D cultured PDLSCs mesenchymal stem cells. The number of mineralized nodules formed by PDLSCs and the expression level of mineralization-related gene ALP OPN mRNA were significantly higher than those of 2D cultured cells (P0.05).
【作者單位】： 南方醫(yī)科大學(xué)南方醫(yī)院;南方醫(yī)科大學(xué)口腔醫(yī)學(xué)院;南方醫(yī)科大學(xué)附屬?gòu)V東省口腔醫(yī)院;
【基金】：國(guó)家自然科學(xué)基金面上項(xiàng)目(81371137)
【分類號(hào)】：R780.2

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本文編號(hào)：1579584