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目的建立CD133+结直肠癌SW480细胞系克隆亚系。方法通过有限稀释法进行单细胞克隆培养,从结直肠癌SW480细胞系48个单细胞孔中培养出8个单细胞源性细胞亚系(编号Y1~8)。对所建立的细胞亚系进行RT-PCR、流式细胞分析及免疫细胞化学筛选CD133+的细胞亚系(Y6、Y2);并通过MTT法体外增殖实验、划痕实验、体外运动实验和皮下成瘤实验,比较CD133+之间及与CD133-单细胞源性细胞亚系的生物学特性。结果筛选出CD133+单细胞源性细胞亚系2个(Y2、Y6),CD133-单细胞源性克隆6个(Y1、Y3、Y4、Y5、Y7、Y8)。结论通过单细胞克隆培养建立CD133+结直肠癌SW480细胞亚系,为下一步结直肠癌转移机制研究提供相关的单细胞源性细胞亚系。
Objective To establish a clonal subline of CD133 + colorectal cancer SW480 cell line. Methods Single cell clone culture was carried out by limiting dilution method, and 8 single cell-derived cell sublines (numbered Y1 ~ 8) were cultured from 48 single-cell wells of colorectal cancer SW480 cell line. The established cell sublines were screened by RT-PCR, flow cytometry and immunocytochemistry for CD133 + cell subline (Y6, Y2). MTT assay was used to detect CD133 + cell proliferation in vitro. Scratch assay, in vitro test and subcutaneous Oncology experiments comparing the biological properties of CD133 + and CD133-monocyte-derived cell sublines. Results Two CD133 + single-cell subline (Y2, Y6) and CD133-single cell clone (Y1, Y3, Y4, Y5, Y7, Y8) were screened. Conclusion Single cell clone culture can establish CD133 + colorectal cancer SW480 cell line and provide relevant single cell lineage for the study of metastasis of colorectal cancer.