Cristofalo V J, Volker C, Allen R G
Center for Gerontological Research, Allegheny University of the Health Sciences, Philadelphia, PA.
Methods Mol Med. 2000;38:23-52. doi: 10.1385/1-59259-070-5:23.
In this chapter, we present standard procedures for the culture of human cells that exhibit a finite proliferative capacity (replicative life-span). The use of a cell culture model has the advantage of providing a controlled environment to study a wide variety of cellular phenomena. It also has the inherent limitation of isolating cells from the regulatory elements that might be provided by other types of cells in vivo. Nevertheless, cell culture models have been crucial to our current understanding of mechanisms of growth, differentiation, development, and neoplasia and numerous other disease states. In this chapter we present procedures for human fibroblast culture including serumfree cultivation of cells, which is necessary when the cellular environment must be fully defined. In addition, we present procedures for the determination of replicative life-span, saturation density, and assessment of replicative capacity from labeled thymidine incorporation in fibroblasts. The methods described here have been well tested and provide highly reproducible results (1, 2).
在本章中，我们介绍了用于培养具有有限增殖能力（复制寿命）的人类细胞的标准程序。使用细胞培养模型的优点是能够提供一个可控环境来研究各种各样的细胞现象。它也有固有的局限性，即把细胞与体内其他类型细胞可能提供的调节元件隔离开来。然而，细胞培养模型对于我们目前对生长、分化、发育、肿瘤形成以及许多其他疾病状态机制的理解至关重要。在本章中，我们介绍人类成纤维细胞培养的程序，包括细胞的无血清培养，当必须完全确定细胞环境时这是必要的。此外，我们还介绍了测定复制寿命、饱和密度以及通过标记胸苷掺入成纤维细胞来评估复制能力的程序。这里描述的方法经过了充分测试，并能提供高度可重复的结果（1, 2）。
