电融合法制备骨肉瘤融合细胞瘤苗的特性分析

Characterization of Osteosarcoma Vaccine Cells Prepared by Electrical Cell Fusion

目的本研究旨在制备骨肉瘤融合细胞瘤苗并检测其体外生物学特性。方法应用电融合技术制备UMR-106细胞与大鼠巨噬细胞的融合细胞,经过磁性分选后,流式细胞仪检测融合细胞的DNA含量和表面抗原的表达。细胞培养检测融合细胞的克隆形成能力,并体外测定其诱导CTL的杀伤活性。结果电融合与磁性分选技术相结合可获得95%的融合细胞纯度。流式细胞仪测定表明,融合细胞可表达较高的MHC-Ⅰ、Ⅱ类抗原。其软琼脂克隆形成能力明显降低(UMR-106,29.2±7.2,融合细胞,17.6±5.1,P<0.05),而诱导CTL细胞杀伤的能力则显著提高(100∶1时UMR-10613.4±3.2,融合细胞42.9±7.5,P<0.05)。结论骨肉瘤融合细胞可稳定生长,在体内诱导CTL细胞产生较强的抗肿瘤活性,可用于动物模型以检测其所诱导的抗骨肉瘤效果。

Objective Prepare the osteosarcoma vaccine by cell fusion and detect itsbiological properties. Methods The osteosarcoma vaccine was obtained by fusing UMR-106 and rat macrophages with electorporator. The DNA and surface antigen expression of fusion cells purified by magnetic cell sorting unit were assayed with flowcytometry. The colony assay of the cells and the analysis of CTL activity induced by fusion cells in vivo were also performed. Results The higher cell fusion efficiency can be obtained with electroporator and about 95 % purity of fusion cells achieved by magnetic cells sorting unit. The fused osteosarcoma cells expressed more MCH- I , H and formed less colony on soft arga. Those cell intensively induced the CTL activity against wild UMR-106 cells as well. Conclusion The proliferation of fused osteosarcoma cells was stable and the high level CTL activity was induced by them. The cells can be used as vaccine in osteosarcoma treatment evaluation on animal models.