中国组织工程研究
中國組織工程研究
중국조직공정연구
Journal of Clinical Rehabilitative Tissue Engineering Research
2014年
6期
841-846
,共6页
干细胞%骨髓干细胞%人骨髓间充质干细胞%神经元样细胞分化%体外实验%表皮生长因子%自噬%Beclin-1%江苏省自然科学基金
榦細胞%骨髓榦細胞%人骨髓間充質榦細胞%神經元樣細胞分化%體外實驗%錶皮生長因子%自噬%Beclin-1%江囌省自然科學基金
간세포%골수간세포%인골수간충질간세포%신경원양세포분화%체외실험%표피생장인자%자서%Beclin-1%강소성자연과학기금
autophagy%celldifferentiation%mesenchymal stem cells%glial fibril ary acidic protein%neurons
背景：骨髓间充质干细胞具有自我更新和多向分化的潜能，但是经过长期体外培养后，骨髓间充质干细胞增殖分化能力逐渐丧失，其机制仍不明确。<br> 目的：观察人骨髓间充质干细胞神经元样细胞分化过程中自噬相关基因表达的变化。<br> 方法：利用表皮生长因子诱导人骨髓间充质干细胞分化为神经元样细胞，观察其形态变化及生长情况。免疫细胞化学法检测神经元特异性烯醇化酶、胶质纤维酸性蛋白表达，采用Western blot检测诱导前后人骨髓间充质干细胞自噬相关基因Beclin-1表达的变化。<br> 结果与结论：经诱导后，人骨髓间充质干细胞呈典型神经元样细胞分化，神经元特异性烯醇化酶阳性率为78.7%，胶质纤维酸性蛋白阳性率为8.1%。诱导0.5 h后处理组细胞中Beclin-1的表达水平有所增加，但1 h后开始逐渐降低。说明体外可以诱导人骨髓间充质干细胞分化为神经元样细胞，自噬相关基因表达活性在诱导分化早期呈高表达，分化过程中逐渐降低。
揹景：骨髓間充質榦細胞具有自我更新和多嚮分化的潛能，但是經過長期體外培養後，骨髓間充質榦細胞增殖分化能力逐漸喪失，其機製仍不明確。<br> 目的：觀察人骨髓間充質榦細胞神經元樣細胞分化過程中自噬相關基因錶達的變化。<br> 方法：利用錶皮生長因子誘導人骨髓間充質榦細胞分化為神經元樣細胞，觀察其形態變化及生長情況。免疫細胞化學法檢測神經元特異性烯醇化酶、膠質纖維痠性蛋白錶達，採用Western blot檢測誘導前後人骨髓間充質榦細胞自噬相關基因Beclin-1錶達的變化。<br> 結果與結論：經誘導後，人骨髓間充質榦細胞呈典型神經元樣細胞分化，神經元特異性烯醇化酶暘性率為78.7%，膠質纖維痠性蛋白暘性率為8.1%。誘導0.5 h後處理組細胞中Beclin-1的錶達水平有所增加，但1 h後開始逐漸降低。說明體外可以誘導人骨髓間充質榦細胞分化為神經元樣細胞，自噬相關基因錶達活性在誘導分化早期呈高錶達，分化過程中逐漸降低。
배경：골수간충질간세포구유자아경신화다향분화적잠능，단시경과장기체외배양후，골수간충질간세포증식분화능력축점상실，기궤제잉불명학。<br> 목적：관찰인골수간충질간세포신경원양세포분화과정중자서상관기인표체적변화。<br> 방법：이용표피생장인자유도인골수간충질간세포분화위신경원양세포，관찰기형태변화급생장정황。면역세포화학법검측신경원특이성희순화매、효질섬유산성단백표체，채용Western blot검측유도전후인골수간충질간세포자서상관기인Beclin-1표체적변화。<br> 결과여결론：경유도후，인골수간충질간세포정전형신경원양세포분화，신경원특이성희순화매양성솔위78.7%，효질섬유산성단백양성솔위8.1%。유도0.5 h후처리조세포중Beclin-1적표체수평유소증가，단1 h후개시축점강저。설명체외가이유도인골수간충질간세포분화위신경원양세포，자서상관기인표체활성재유도분화조기정고표체，분화과정중축점강저。
BACKGROUND:Bone marrow mesenchymal stem cells have potential to self-renewal and multi-lineage differentiation. But after a long period of culture in vitro, the proliferation and differentiation capacities of bone marrow mesenchymal stem cells gradual y loss, the mechanism underlying which is not clear now. <br> OBJECTIVE:To observe the expression of autophagy-related gene Beclin-1 in differentiation from human bone marrow mesenchymal stem cells into neuron-like cells in vitro. <br> METHODS:The changes of morphological characteristics of neuron-like cells differentiated from human bone marrow mesenchymal stem cells induced by epidermal growth factor were observed. The expression of neuron-specific enolase and glial fibril ary acidic protein in treated and untreated human bone marrow mesenchymal stem cells were detected using immunocytochemistry. The Beclin-1 protein expressions were detected by western blot before and after induction. <br> RESULTS AND CONCLUSION:After being induced, human bone marrow mesenchymal stem cells presented classical neuron-like morphology;the expressions of neuron-specific enolase and glial fibril ary acidic protein were 78.7%and 8.1%, respectively. The expression of Beclin-1 protein was changed correspondingly during the induction, which increased after 30 minutes of induction and decreased gradual y after 1 hour of induction. Human bone marrow mesenchymal stem cells could be induced into neuron-like cells in vitro by epidermal growth factor. Autophagy-related gene was highly expressed in the induction of early differentiation and the expression gradual y reduced until it remained at a low level during the differentiation.
