Effect of Serum Media on Fibroblast Proliferation and Collagen Synthesis

Lee, Min Ah; Seo, Sung Ig; Han, Seung Kyu; Kim, Woo Kyung

Abstract

Expanding cells ex-vivo is very important in tissue- engineering. Culture medium is usually supplemented with fetal bovine serum(FBS) in most of the experiments. However, cells grown in bovine serum media may posses the possibilities of disseminating bovine diseases and/or stimulating the patient's immune reactions. To overcome these problems, autologous or homologous serum should be used instead of the FBS. The purpose of this study is to compare cell proliferation and collagen synthesis depending on the kind of sera mixed on media and to provide a guideline on applying established experimental data to clinical cases. Human dermal fibroblasts were obtained from four patients. Five thousand cells per well in 96-well plates were incubated DMEM/F- 12 Nutrient with varying serum mixture; 10% autologous serum, 10% homologous serum, and 10% FBS. Five days after incubation fibroblast proliferation and collagen production were determined by MTT assay and CICP enzyme immunoassay. The mean cell number were; 3.95x104/well, 2.97x104/well and 2.30x104/well, respectively. The average amounts of collagen synthesized were; 238.13ng/ml, 204.88ng/ml, and 163.88ng/ml in each. These results show that the use of human serum mixture may contribute to, not only preventing disseminated infection of bovine diseases. but also increase cell proliferation and collagen synthesis without simulating the patient's immune reactions.