Objectives This study was carried out to investigate the effect of Ganoderma lucidum extract (GLE) and microneedle therapy system (MTS) on hair growth in an alopecia model of C57BL/6N mice. Methods Five-week old mice were depilated and separated in 4 groups ; CON (50% EtOH), MXD (5% Minoxidil), MTS and GLE + MTS. The treatments were applied twice a week for 3 weeks. The hair growth was determined photographically, the hair density, thickness and length were identified by Folliscope and the weights of body and organs were measured. In dorsal skin tissue, the expression of hair growth-related gene and protein was analyzed by RT - PCR or Western blot. In addition, the hair follicles in the dermis were observed by H&E staining. Results The promotion of hair growth was observed in GLE + MTS and MTS compared to CON. The hair density, thickness and length were also improved in GLE + MTS and MTS compared to CON. The mRNA expression of TGF-${alpha}$, TGF-${eta}1$, IGF-1, PRL and PL and the protein expression of VEGF and IGF-1 were increased in GLE + MTS and MTS compared to CON. The hair follicles and hair root growth were improved in GLE + MTS and MTS compared to CON. In the above results, GLE + MTS were more effective than MTS. Conclusions These results suggest that GLE and MTS has a hair growth activity and can be useful for the treatment of alopecia.