Effect of culture substrate and fibroblast growth factor addition on the proliferation and differentiation of human adipo-stromal cells

Abstract

The objective of this study is to investigate the proliferation and differentiation of stromal cells derived from human adipose tissues cultured on substrates with different surface properties. In addition, a similar investigation was performed on cells proliferated in different concentrations of basic fibroblast growth factor (FGF-2). The culture substrates include several polymer films with different water wettabilities, glass or a cell-culture plate, and that coated with collagen type I or IV, gelatin and FGF-2. The proliferation profiles of cells were influenced by the type of culture substrate and the growth factor concentration. A larger number of proliferated cells was observed for substrates with a water contact angle around 80°, while the cell number was significantly larger for every protein-coated substrate. The rate of cell proliferation became maximal at a FGF-2 concentration of 1000 ng/ml. The FGF-2 concentration used for cell proliferation affected the differentiation profile of cells proliferated. Stromal cells, proliferated in 1 ng/ml FGF-2, were osteogenically differentiated to the strongest and fastest extent among those in other growth factor doses. The alkaline phosphatase (ALP) activity of cells increased with the increased cell number, although the activity per cell was identical, irrespective of the substrate type. The strongest adipogenic differentiation was observed for cells proliferated in 1000 ng/ml FGF-2 and the differentiation induction was maintained for a long time period. No clear dependence of the cell number on adipogenesis was observed. These findings indicate that the proliferation and differentiation of human adipose tissue-derived stromal cells are influenced by the culture substrate and the concentration of FGF-2 used for proliferation.