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目的:探讨大鼠成肌细胞体外能否转化为成骨前体细胞。方法:从新生大鼠骨骼肌分离单细胞,体外培养,运用差速贴壁法和(或)酶消化法纯化细胞,培养至第3代时,加入含重组人骨形态发生蛋白-2(rhBMP-2)的培养液进行诱导,3周后对诱导细胞进行鉴定,其指标包括:细胞形态、碱性磷酸酶染色及其活性、Ⅰ型胶原免疫组化染色和钙结节染色,实验数据采用SAS9.1.3软件包进行统计学分析。结果:成肌细胞诱导后,细胞增殖减缓,相邻细胞聚集呈层状排列,诱导7d可见胞浆有少量不透光结节,诱导2周结节增多,3周时胞浆内可见大量不透光结节,对照组成肌细胞融合成有收缩性的肌管。诱导后的成肌细胞碱性磷酸酶活性随时间延长而增强,诱导3周时碱性磷酸酶染色、Ⅰ型胶原和钙结节染色均呈阳性表达。结论:大鼠成肌细胞在体外一定条件下诱导后能够转化为成骨前体细胞。
Objective: To investigate whether in vitro rat myoblasts can be transformed into osteoblast precursors. Methods: Single cells were isolated from skeletal muscle of newborn rats and cultured in vitro. Cells were purified by differential adherent method and / or enzyme digestion method. When cultured to the third passage, rhBMP- 2). After 3 weeks, the induced cells were identified. The indexes included cell morphology, alkaline phosphatase staining and its activity, type I collagen immunohistochemistry staining and calcium nodule staining. The experimental data were analyzed by SAS9 .1.3 software packages for statistical analysis. Results: After induction of myoblasts, the proliferation of cells slowed down and the adjacent cells gathered in a layered arrangement. A small amount of opaque nodules were observed in the cytoplasm at 7 days after induction, and the number of nodules increased at 2 weeks. Translucent nodules, the control group myogenic cells fused into contractile myotubes. After induction, the alkaline phosphatase activity of myoblasts increased with time. Alkaline phosphatase staining, type I collagen and calcium nodule staining were all positive after induction for 3 weeks. CONCLUSION: Rat myoblasts can be transformed into osteoblast precursors after induction under certain conditions in vitro.