本文選題：牙周膜干細胞　切入點：D培養(yǎng)　出處：《牙體牙髓牙周病學雜志》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)的牙周膜干細胞(PDLSCs)生物學特性的差異。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)按不同細胞接種數分為2.5×104,5×104及1×1053組),培養(yǎng)96 h后分別用Live/Dead染色法分析細胞活性;流式細胞術檢測干細胞表面標記物;然后對2D、3D培養(yǎng)的PDLSCs進行礦化誘導,茜素紅染色及熒光定量PCR檢測礦化結節(jié)形成情況以及礦化相關基因ALP和OPN mRNA的表達水平。結果:3D培養(yǎng)的PDLSCs可形成致密的細胞聚合物,2.5×104組PDLSCs細胞聚合物中心未見壞死現象,5×104組和1×105組則有較為明顯壞死現象;3D和2D培養(yǎng)的PDLSCs間充質干細胞表面標記物的陽性表達率均無顯著差異(P0.05);茜素紅染色和熒光實時定量PCR結果顯示,3D培養(yǎng)的PDLSCs形成礦化結節(jié)的數量及其礦化相關基因ALP OPN mRNA的表達水平均顯著高于2D培養(yǎng)的細胞(P0.05)。結論: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)科大學南方醫(yī)院;南方醫(yī)科大學口腔醫(yī)學院;南方醫(yī)科大學附屬廣東省口腔醫(yī)院;
【基金】：國家自然科學基金面上項目(81371137)
【分類號】：R780.2

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