Abstract: Objective To investigate the possibility of culturing human scalp skin in serum-free medium, and to observe the changes in the function and structure of scalp skin. Methods Intact human scalp skin was obtained from plastic surgery, and cultured at the air/liquid interface in serum-free William′s E medium supplemented with penicillin, streptomycin, hydrocortisone, etc. MEM supplemented with serum served as control. Dissection microscopy was used to observe the growth of hairs. The changes of scalp structure were examined by hematoxylin and eosin staining, and lactate dehydrogenase （LDH） activity by cytotoxicity detection kit. Results The human scalp skin maintained an intact structure after a 22-day organ-culture in vitro in William′s E medium. The hair growth was more rapid in serun-free William′s E medium compared with serum-containing MEM （P < 0.05）. LDH activity experienced 2 peaks on day 2 and 16 after culturing in serun-free William′s E medium, but only 1 peak on day 12 in serum-containing MEM. With the prolongation of culture period, hair follicles gradually turned from anagen to catagen and telogen. Conclusion The organ culture system may serve as a valuable tool for scalp skin and hair research.