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目的比较观察人肝癌细胞株BEL7402细胞与LAK细胞体外凋亡的超微结构.方法将BEL7402细胞、人血树突状细胞和人LAK细胞(淋巴因子和PHA激活的杀伤细胞)共同置于含100mL/L新生牛血清的1640培养液内,在37℃,50mL/L,CO2,饱湿条件下培养6h.另将LAK细胞单独置于含有环磷酰胺的上述培养液内,在同样条件下培养6h.收集沉淀细胞制备电镜样本.细胞经25g/L戊二醛和20g/L锇酸双固定,PDAP包埋,超薄切片,常规染色,透射电镜观察.结果凋亡的BEL7402细胞与LAK细胞,其细胞核染色质浓缩、边集,核碎裂,离散的碎片外包以双层膜,一些线粒体也变得致密.然而,在有些凋亡的BEL7402细胞内,可见浓缩的细胞碎片向细胞外出芽隆起,形成膜包的凋亡小体;其线粒体和粗面内质网的变化也较为复杂.结论凋亡的BEL7402细胞与LAK细胞均具有凋亡细胞的基本特征,但是两种凋亡细胞具体的形态表现不尽一致.表明细胞内的主要细胞器主动参与了凋亡过程.
4. Objective To observe the ultrastructure of apoptosis of human hepatocellular carcinoma cell line BEL7402 and LAK in vitro. Methods BEL7402 cells, human blood dendritic cells and human LAK cells (lymphokine and PHA-activated killer cells) were co-cultured in 1640 medium containing 100 mL/L newborn calf serum at 37°C, 50 mL/L, CO2. , Cultivation under humid conditions 6h. The LAK cells were separately placed in the culture medium containing cyclophosphamide and cultured for 6 h under the same conditions. Precipitated cells were collected for electron microscopy. The cells were fixed with 25g/L glutaraldehyde and 20g/L tannic acid, embedded in PDAP, ultrathin sections, routine staining, and transmission electron microscope observation. RESULTS: Apoptotic BEL7402 cells and LAK cells, whose nuclear chromatin was concentrated, edged, and nuclear fragmented, the discrete fragments were overwhelmed with double membranes, and some mitochondria became dense. However, in some of the apoptotic BEL7402 cells, concentrated cell debris can be seen to sprout out of the cell to form apoptotic bodies, and the changes of mitochondria and rough endoplasmic reticulum are more complex. Conclusion The apoptotic BEL7402 cells and LAK cells both have the basic features of apoptotic cells, but the specific morphology of the two apoptotic cells is not consistent. It indicates that the main organelle in the cell actively participates in the apoptosis process.