Objective  To establish a simple stirring-migration culture system for human dermal fibroblasts (HDFs) by stirring in the three-dimensional cultivation monitored by electron microscope and to research its migration primarily.   Methods  Skin-derived primary HDFs were obtained by enzymatic digestion of foreskin

and then the cells at passage 5 to 8 were further cultured in the presence of microbeads with stirring for in vitro amplification. Static culture was also set up as control. The morphology of HDFs on the microcarriers in the stirring culture system was observed with inverted phase contrast microscopy and scanning electron microscopy (SEM). Then the cells were migrated from the attached microcarriers to tissue culture flasks. The cells were observed in every 24 h by inverted phase contrast microscopy. HDFs phenotype was identified with immunocytochemical staining against collagen Ⅰ and vimentin

and ELISA for the content of collagenⅠ in the supernatant of migration culture indirectly.   Results  HDFs adhered to microcarriers quickly with normal appearance. The morphology of HDFs did not show significant changes when compared with HDFs cultured in normal static culture. HDFs automatically and quickly migrated from microcarriers to the bottom surface of culture flask firmly. HDFs showed fusiform or polygonal appearance and their growth patterns showed blossom or whirlpool. So the morphology and growth pattern of these migrated cells were normal

in addition to the expression of collagen I and vimentin

and collagen Ⅰ secretion in accordance with the cells’number.   Conclusion  The established stirringmigration culture of HDFs is a simple method for quick amplification of HDFs in vitro

which may meet the demand for various experiments or production of seed cells in tissue engineering.