Simonian M H, White M L, Foggia D A
In Vitro Cell Dev Biol. 1987 Apr;23(4):247-52. doi: 10.1007/BF02623705.
The life span and growth from clonal density of bovine adrenocortical cell cultures were studied in serum-supplemented medium and a serum-free defined medium, which supported sustained cell proliferation and steroid production. The total culture life span was 79 population doublings in serum-supplemented medium with fibroblast growth factor (FGF) and 36 population doublings in the defined medium without serum. Older passage cell cultures grown in the defined medium progressively lost the ability to produce 11 beta- and 21-hydroxylated steroids, which was observed previously for cultures in serum-supplemented medium, and also had a decline of 17 alpha-hydroxylated steroid production. The cloning efficiency in the defined medium was 12.2% as compared to 24% in serum-supplemented medium with FGF. Five isolated clonal cell lines grown in the defined medium were characterized for steroid function in response to steroidogenic agents. All five clonal cell lines had stimulated steroid production with 8-bromo-cAMP, but only four of the clonal lines were stimulated also by adrenocorticotropin. None of the clonal cell lines produced 11 beta-, 21- or 17 alpha-hydroxylated steroids in response to treatment with either steroidogenic agent, results that were similar to data obtained from older mass cultures. The apparent deficiency of the defined medium as compared to serum-supplemented medium for maximum support of the culture life span and cloning efficiency may be useful in studies of cellular aging and its relation to differentiated function for this cell culture system.
在添加血清的培养基和无血清限定培养基中研究了牛肾上腺皮质细胞培养物的寿命以及克隆密度下的生长情况，这两种培养基均能支持细胞持续增殖和类固醇生成。在添加血清并含有成纤维细胞生长因子（FGF）的培养基中，培养物的总寿命为79个群体倍增；在无血清的限定培养基中为36个群体倍增。在限定培养基中培养的传代次数较多的细胞培养物逐渐丧失产生11β-和21-羟基化类固醇的能力，这在之前添加血清的培养基培养物中也有观察到，同时17α-羟基化类固醇的产生也有所下降。限定培养基中的克隆效率为12.2%，而添加血清并含有FGF的培养基中为24%。对在限定培养基中生长的5个分离的克隆细胞系进行了针对类固醇生成剂的类固醇功能特性分析。所有5个克隆细胞系用8-溴-cAMP刺激后类固醇生成均增加，但只有4个克隆系也受到促肾上腺皮质激素的刺激。在用任何一种类固醇生成剂处理后，没有一个克隆细胞系产生11β-、21-或17α-羟基化类固醇，这些结果与从传代次数较多的大量培养物中获得的数据相似。与添加血清的培养基相比，限定培养基在最大程度支持培养物寿命和克隆效率方面明显不足，这对于该细胞培养系统的细胞衰老及其与分化功能关系的研究可能有用。
