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目的 探索从骨肉瘤细胞条件培养基中提取骨形态蛋白(BMP)的方法,并测定其生物学活性。方法 收集骨肉瘤细胞(MG-63)条件培养基,通过浓缩、透析,SephcrylS-100凝胶层析纯化,BMP单克隆抗体鉴定所需洗脱峰,SDS-PAGE测定分子量,小鼠肌袋实验检测其骨诱导活性。结果 BMP单抗鉴定所提蛋白为BMP,SDS-PAGE显示分子量在21kD,能够在小鼠肌肉内产生异位骨化。结论 骨肉瘤细胞条件培养基中含有BMP,分离后具有良好的生物学活性,而骨肉瘤细胞可以在体外长期培养生长,为BMP的大量提取、临床应用提供一个有益的方法。
Objective To explore the method of extracting bone morphogenetic protein (BMP) from conditioned medium of osteosarcoma cells and determine its biological activity. METHODS: Osteosarcoma cells (MG-63) conditioned media were collected and purified by concentration, dialysis and Sephcryl S-100 gel chromatography. The desired peak was identified by BMP monoclonal antibody. The molecular weight was determined by SDS-PAGE. The osteoinductive activity was examined. Results BMP monoclonal antibody identified the protein as BMP, SDS-PAGE showed a molecular weight of 21kD, can produce heterotopic ossification in mouse muscle. Conclusion Osteosarcoma cells contain BMP in conditioned medium, which has good biological activity after isolation. However, osteosarcoma cells can be cultured and grown in vitro for a long time. It provides a useful method for extracting large amounts of BMP and clinical application.