Culture and expansion of human dermal fibroblasts using rotary cell culture system

Objective: To study the feasibility of culture and expansion of human dermal fibroblasts(hDFBs) on a large scale by Rotary Cell Culture System(RCCS).Methods: hDFBs were cultured in vitro and labeled with fluorescent dye DIO,then the cells were transfered into RCCS containing culture medium and microcarrier Cytodex-3 at 5 g/L.The characteristics of fibroblasts on microcarriers were observed with fluorescent microscope,and scanning electronic microscope(SEM).Cell cycle and doubling time of RCCS cultured hDFBs were studied by cell counting and flow Cytometry and compared with that of conveitional plate cultured hDFBs(control).Results: In RCCS culture hDFBs attached to Cytodex-3 and proliferated to heigh cell density.The doubling time(d) of RCCS cultured cells was 2.99±0.5,that of controls 3.34±0.42(P0.05).The proliferation index of RCCS group was 48.2±4.3,that of control 35.2±3.4(P0.05).Conclusion: A culture system of biological reactor containing microcarriers is effective in preparation a large number of human skin fibroblasts.