本文選題：骨髓基質(zhì)干細(xì)胞 + 分化　； 參考：《解剖學(xué)報》2016年01期

【摘要】：目的探討體外誘導(dǎo)人骨髓基質(zhì)干細(xì)胞(HMSCs)向角膜上皮樣細(xì)胞分化的可能性。方法組織塊培養(yǎng)法原代培養(yǎng)人眼角膜基質(zhì)細(xì)胞(HCFs)并進行傳代培養(yǎng);密度梯度離心法分離、培養(yǎng)HMSCs;倒置相差顯微鏡觀察細(xì)胞的形態(tài)學(xué)特征;免疫細(xì)胞化學(xué)對細(xì)胞進行鑒定。利用角膜上皮細(xì)胞生長的微環(huán)境對處于共培養(yǎng)體系的HMSCs進行誘導(dǎo)培養(yǎng);免疫細(xì)胞化學(xué)檢測角膜上皮細(xì)胞的特異性分子標(biāo)志角蛋白12(CK12)在分化細(xì)胞的原位表達;反轉(zhuǎn)錄聚合酶鏈反應(yīng)(RT-PCR)檢測分化細(xì)胞CK12 m RNA的表達,用單獨培養(yǎng)的HMSCs做對照。結(jié)果HMSCs和HCFs貼壁生長,以梭形為主,接近匯合的細(xì)胞排列有一定方向性,呈漩渦狀;HMSCs陽性表達CD29;HCFs陽性表達波形纖維蛋白;隨著共培養(yǎng)時間的延長,HMSCs形狀逐漸由長梭形變?yōu)椴灰?guī)則多邊形,與上皮細(xì)胞的形態(tài)相似;免疫細(xì)胞化學(xué)和RT-PCR分別在蛋白和基因水平證實分化細(xì)胞表達CK12。結(jié)論在一定誘導(dǎo)條件下,體外培養(yǎng)的HMSCs可向具有角膜上皮細(xì)胞表型的上皮樣細(xì)胞分化。
[Abstract]:Objective to investigate the possibility of inducing the differentiation of human bone marrow stromal cells (HMSCs) into corneal epithelioid cells in vitro. Methods Human corneal stromal cells (HCFs) were cultured with tissue mass culture method, HMSCs were isolated by density gradient centrifugation, morphological characteristics of cells were observed by inverted phase contrast microscope, and cells were identified by immunocytochemistry. The HMSCs in the co-culture system was induced by the microenvironment of corneal epithelial cell growth, and the expression of keratin 12 (CK12), a specific molecular marker of corneal epithelial cells, was detected by immunocytochemistry. Reverse transcriptase polymerase chain reaction (RT PCR) was used to detect the expression of CK12 m RNA in differentiated cells. HMSCs cultured alone was used as control. Results the adherent cells of HMSCs and HCFs were mainly fusiform, and the cells close to confluence were arranged in a certain direction, and the positive expression of CD29HCFs and fibrin was observed in the form of whirlpool. With the extension of co-culture time, the shape of HMSCs gradually changed from long spindle to irregular polygon, similar to the morphology of epithelial cells. Immunocytochemistry and RT-PCR confirmed the expression of CK12 in differentiated cells at the protein and gene levels, respectively. Conclusion under certain induction conditions, cultured HMSCs can differentiate into epithelioid cells with phenotype of corneal epithelial cells.
【作者單位】： 山東大學(xué)第二醫(yī)院眼科;山東省腫瘤醫(yī)院基礎(chǔ)實驗室;山東大學(xué)醫(yī)學(xué)院人體解剖學(xué)研究所;
【基金】：山東省科技發(fā)展計劃(2014GSF118006) 山東大學(xué)基本科研業(yè)務(wù)費資助自然科學(xué)專項(自由探索項目2015TS012)
【分類號】：R772

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