Factors controlling the proliferative rate, final cell density, and life span of bovine vascular smooth muscle cells in culture.
Factors controlling the proliferative rate, final cell density, and life span of bovine vascular smooth muscle cells in culture.
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DOI：
10.1083/jcb.89.3.568
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发表时间：
1981-06
期刊：
影响因子：
--
通讯作者：
Tauber JP
中科院分区：
文献类型：
--
作者：
Gospodarowicz D;Hirabayashi K;Giguère L;Tauber JP
低密度血管平滑肌（VSM）细胞培养物保持在细胞外基质（ECM）包被的培养皿上，并在血浆或血清存在下铺板，当含血清培养基被补充有三种因子的合成培养基取代时，细胞增殖活跃：高密度脂蛋白5微克/ml）或生长调节素C（10 ng/ml）;和成纤维细胞生长因子（FGF，100 ng/ml）或表皮生长因子（EGF，50 ng/ml）。从合成培养基中省略这三个因素中的任何一个都会导致培养物的生长速率较低，以及一旦培养物达到汇合，最终细胞密度较低。当细胞在完全不存在血清的情况下铺板时，还需要转铁蛋白（10微克/毫升）来诱导最佳细胞生长。还分析了底物和培养基补充剂对VSM培养物寿命的影响。将培养物保持在塑料上并暴露于补充有5%牛血清的培养基中，进行了15代。然而，当维持在ECM包被的培养皿上时，血清喂养的培养物具有至少88代的寿命。同样，当培养物保持在补充有HDL和FGF或EGF的合成培养基中时，观察到底物对组织培养寿命的影响。培养物保持在塑料经历了24代，而那些保持在ECM包被的培养皿可以反复传代58代。这些实验证明了ECM底物不仅在促进细胞生长方面而且在增加培养物寿命方面的影响。
Low density vascular smooth muscle (VSM) cell cultures maintained on extracellular-matrix(ECM)-coated dishes and plated in the presence of either plasma or serum will proliferate actively when serum-containing medium is replaced by a synthetic medium supplemented with three factors: high density lipoprotein (HDL, 250 micrograms protein/ml); insulin (2.5 micrograms/ml) or somatomedin C (10 ng/ml); and fibroblast growth factor (FGF, 100 ng/ml) or epidermal growth factor (EGF, 50 ng/ml). The omission of any of these three factors from the synthetic medium results in a lower growth rate of the cultures, as well as in a lower final cell density once cultures reach confluence. When cells are plated in the total absence of serum, transferrin (10 micrograms/ml) is also required to induce optimal cell growth. The effects of the substrate and medium supplements on the life span of VSM cultures have also been analyzed. Cultures maintained on plastic and exposed to medium supplemented with 5% bovine serum underwent 15 generations. However, when maintained on ECM-coated dishes the serum-fed cultures had a life span of at least 88 generations. Likewise, when cultures were maintained in a synthetic medium supplemented with HDL and either FGF or EGF, an effect on the tissue culture life span by the substrate was observed. Cultures maintained on plastic underwent 24 generations, whereas those maintained on ECM-coated dishes could be passaged repeatedly for 58 generations. These experiments demonstrate the influence of the ECM-substrate only in promoting cell growth but also in increasing the longevity of the cultures.