A REPRODUCIBLE PROCEDURE FOR PRIMARY CULTURE AND SUBSEQUENT MAINTENANCE OF MULTIPLE LINES OF HUMAN SKIN FIBROBLASTS

Cultured fibroblasts are a valuable tool to study many cellular proces ses and their modification by aging. Fibroblasts are a useful cell typ e in which to study many diseases, including those of the nervous syst em, in which a strong genetic component is suspected. Fibroblasts perm it the study of multiple, dynamic processes in living cells, while avo iding the effects of the dying process and post-mortem artifacts that limit other approaches. For results to be comparable across time in on e laboratory or consistent between laboratories, the detailed culture techniques require meticulous care and replicability. Lack of attentio n to detail in initial stages can lead to selection of different cell populations. Small variations in other variables such as batches of se rum can significantly alter growth rates and comparisons of cells from controls and Alzheimer patients. The aim of this paper is to present a detailed protocol for comparison of multiple cell lines from many pa tients. An example of using this approach to study growth and phase ou t (i.e., senescence) of cells from Alzheimer patients is presented. Th is procedure represents a modification of an earlier protocol (Cristof alo and Charpentier, 1980).