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wangshixiaowsx金虫 (小有名气)
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目的:探讨A的含药血清体外诱导大鼠骨髓基质干细胞分化为心肌样细胞的可行性。 方法:用DMEM/F-12培养液培养Wistar大鼠骨髓基质干细胞(BMSCs),采用台盼兰排斥试验检测BMSCs活力。用A的含药血清诱导BMSCs,采用酶联免疫检测其OD值。诱导分化后,倒置显微镜观察细胞形态变化;用免疫细胞化学染色法、免疫细胞荧光染色法以及PCR技术鉴定表达。 结果:诱导7天后,骨髓基质干细胞细胞变宽,呈长梭形、圆形和椭圆形并伸出细长突起,在突起的末端出现分支,排列方向渐趋一致;诱导4周后,部分相邻细胞的突起连接成网,可见肌管样结构。免疫细胞化学染色示结蛋白(Desmin),肌动蛋白(actin)和心肌肌钙蛋白(cTnI)的表达阳性表达;荧光显微镜示心肌肌钙蛋白(cTnI)阳性表达;利用PCR法检测诱导后的细胞中有心肌细胞标志物α-MHC,β-MHC的表达。 结论:A的含药血清可以体外诱导大鼠骨髓基质干细胞分化为心肌样细胞。 |
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【答案】应助回帖
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Mally89: 金币+1, 感谢应助!~欢迎常来!~吼吼!~\(^o^)/~ 2012-03-19 13:36:27
wangshixiaowsx: 金币+40, 翻译EPI+1, ★有帮助 2012-03-20 22:14:16
sltmac: 金币+20 2012-03-26 12:32:32
Mally89: 金币+1, 感谢应助!~欢迎常来!~吼吼!~\(^o^)/~ 2012-03-19 13:36:27
wangshixiaowsx: 金币+40, 翻译EPI+1, ★有帮助 2012-03-20 22:14:16
sltmac: 金币+20 2012-03-26 12:32:32
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Purpose: In this paper, we aim to discuss feasibility on how serum containing in A externally change the Rat Bone Marrow Stromal Stem Cells into Cardia-lick Cells. Method: In this research, Wistar BMSCS can be produeced by DMEM/F-12 culture medium. Meanwhile, we detected the activity of BMSCS through trypan blue exclusion testing. Serum containing in A induces BMSCS as well as tesing OD value by enzyme linked immunosorbent . Morphological changes can be found by using inverted microscope, after differentiation. Then, It may be indicated by immunocytochemical staining, and immunoc- ytological fluorescent staining, PCR. Conclusion: MBSCs is wider, showing long fusiform, circle and Elliptic after 7 days inducing. Little branch can be found in the end of emerged parts with same direction; The adjacent cells form a net rosily, which indicates the myotube-like cells structure; Further, Immunocytochemical staining show the positive expression by using Desmin, Actin, CTNI. Fluorescence microscope express CTnI positively. By using PCR, α-MHC,β-MHC in cardiac cell can be obtained. In conclusion, differentiation from BMSCs in Vitro to cardiac-like cells by using Serum Containing in A is feasible. |
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