Abstract: Objective To prepare a composit membrane with collagen scaffold material for tissue culture. Methods Pure collagen membrane, collagen/hyaluronic acid (HA) membrane, collagen/chitosan membrane and collagen/chitosan/HA membrane (abbreviated as composite collagen membrane) were prepared respectively, and their respective biodegradation time and the number of the cells proliferated on them were examined in vitro. Result The biodegradation time of pure collagen, collagen/HA and composite collagen membranes in vitro was 1380, 1376 and 1560 min respectively, and the number of cells proliferated on the collagen/HA membrane and the composite collagen membrane were greater than that on the pure collagen membrane. Conclusions Addition of chitosan can decrease the biodegradation of the collagen membrane in vitro, while HA can not; HA Can contribute to cell adhesion and proliferation on the collagen membranes, but chitosan does not.