Control of cytolysis of BALB/c-3T3 cells by platelet-derived growth factor: a model system for analyzing cell death.
Control of cytolysis of BALB/c-3T3 cells by platelet-derived growth factor: a model system for analyzing cell death.
复制标题
血小板衍生生长因子控制 BALB/c-3T3 细胞的细胞溶解：分析细胞死亡的模型系统。
DOI：
10.1002/jcp.1041130205
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发表时间：
1982
影响因子：
5.6
通讯作者：
Locatell,KL
中科院分区：
文献类型：
--
作者：
Scher,CD;Young,SA;Locatell,KL
在添加10%凝血血清的培养基中，通过细胞复制和细胞损失共同确定BALB/c‐3T3细胞的饱和密度。通过用3h‐胸苷预标记细胞DNA和延时摄影，我们研究了独立于复制的细胞损失。当BALB/c‐3T3细胞从含有血小板衍生生长因子（PDGF）的补充血清培养基转移到缺乏血小板衍生生长因子的补充血浆培养基时，细胞损失加速。细胞通过附着在组织培养皿表面的细胞解体而丧失。单个细胞迅速溶解；在第一个摄动迹象（通过相衬显微镜）和细胞质破裂之间不到15分钟。动力学分析与随机细胞死亡一致，而不是固定寿命。细胞溶解的百分比由细胞密度决定；在高密度下，如在融合培养中，细胞损失的百分比比在低密度下高。部分纯化或电泳均相的PDGF制剂可拮抗细胞死亡。纯PDGF以ng/ml浓度刺激细胞存活，呈浓度依赖性。细胞复制过程对于存活来说不是必需的，因为PDGF在DNA合成抑制剂甲氨蝶呤的存在下阻止了细胞溶解。PDGF短暂（4小时）治疗可防止细胞死亡；这种PDGF处理的细胞在胰蛋白酶处理后，在补充血浆的培养基中移植到新鲜表面，显示出更高的存活率。垂体成纤维细胞生长因子（垂体成纤维细胞生长因子）是PDGF在BALB/c‐3T3细胞中诱导DNA合成的功能类似物，也具有抗溶细胞剂的功能。相比之下，表皮生长因子和胰岛素则没有。SV40转化细胞的细胞溶解率较低，对PDGF不敏感。
In culture medium supplemented with 10% clotted blood serum, the saturation density of BALB/c‐3T3 cells is determined jointly by cell replication and cell loss. By prelabelling cellular DNA with3H‐thymidine and also by time lapse photography, we studied cell loss independently of replication. Cell loss was accelerated when BALB/c‐3T3 cells were transferred from serum‐supplemented medium, which contains the platelet derived growth factor (PDGF), to medium supplemented with platelet‐poor plasma which lacks it. Loss occurred via the disintegration of cell attached to the surface of the tissue culture dish. Cytolysis of individual cells occurred rapidly; less than 15 minutes transpired between the first indication of a perturbance (by phase contrast microscopy) and fragmentation of the cell cytoplasm. Kinetic analysis was consistent with random cell death rather than a fixed lifetime.The percentage of cells undergoing cytolysis was governed by the cell density; at high densities, such as are present in confluent cultures, a higher percentage of cell loss was noted than at low density. Cell death was antagonized by partially purified or electrophoretically homogenous preparations of‐PDGF. Pure PDGF stimulated cell survivial at ng/ml in a concentration dependent fashion. The process of cell replication was not necessary for survival because PDGF prevented cytolysis in the presence of methotrexate, an inhibitor of DNA synthesis. A brief (4 hour) treatment with PDGF prevented cell death; such PDGF treated cells displayed increased survival after being taken up with trypsin and planted onto a fresh surface in plasma supplemented medium. Pituitary fibroblast growth factor, a functional analogue of PDGF for induc of DNA synthesis in BALB/c‐3T3 cells, also functioned as an anticytolytic agent. By contrast, epidermal growth factor and insulin did not. Cytolysis of SV40‐transformed cells occurred at a constitutively low rate and was insensitive to PDGF.