Xiong Y M, Xu S Z, Slakey L L
Department of Biochemistry, University of Massachusetts, Amherst 01003.
In Vitro Cell Dev Biol. 1991 May;27A(5):355-62. doi: 10.1007/BF02630954.
Cultured pig aortic smooth muscle cells maintain a viable, quiescent state in a chemically defined medium that contains 10(-6) M insulin, 5 micrograms/ml transferrin, and 0.2 mM ascorbate. DNA synthesis and DNA content were determined by measuring tritiated thymidine incorporation and DNA-binding to the fluorescent probe 4',6-diamidino-2-phenylindole, respectively. The majority of the population of cells in defined medium cultures were diploid. Tritiated thymidine uptake in cells in defined medium was one-tenth that observed in cells in fetal bovine serum-containing medium. The study of cellular cyclic AMP level in response to extracellular adenosine stimulation in dividing cells and quiescent cells showed that cells in defined medium had a lower extent of response to adenosine compared to cells cultured in serum-containing medium. Both the cell growth index and the response to adenosine of cells cultured in defined medium were reversible after replacing the medium with 10% fetal bovine serum-containing medium, which suggests that the cells in defined medium were healthy and were capable of modulating cellular metabolism depending on culture conditions.
培养的猪主动脉平滑肌细胞在含有10⁻⁶ M胰岛素、5微克/毫升转铁蛋白和0.2 mM抗坏血酸盐的化学成分确定的培养基中维持存活的静止状态。分别通过测量氚标记胸腺嘧啶核苷掺入和DNA与荧光探针4',6-二脒基-2-苯基吲哚的结合来测定DNA合成和DNA含量。在化学成分确定的培养基中培养的大多数细胞群体是二倍体。在化学成分确定的培养基中细胞对氚标记胸腺嘧啶核苷的摄取是在含胎牛血清培养基中细胞摄取量的十分之一。对分裂细胞和静止细胞中细胞环磷酸腺苷水平对细胞外腺苷刺激的反应的研究表明，与在含血清培养基中培养的细胞相比，化学成分确定的培养基中的细胞对腺苷的反应程度较低。在用含10%胎牛血清的培养基替换培养基后，化学成分确定的培养基中培养的细胞的细胞生长指数和对腺苷的反应都是可逆的，这表明化学成分确定的培养基中的细胞是健康的，并且能够根据培养条件调节细胞代谢。
