Background: Growth factor cocktail (GFC) treatment with microneedling in androgenetic alopecia (AGA) patients is effective and safe. However, there is a lack of study on its effect including fibroblast growth factor 9 (FGF9). Objectives: Importance of FGF9 in hair follicle regeneration was reported. The aim was to evaluate the efficacy of GFC including FGF9 on AGA. Methods: The scalp was divided to right and left and treated with GFC including FGF9 and normal saline, respectively; both sides were topically applied by medical devices of microneedle once in 2 weeks for 3 months. 9 males and 9 females were enrolled. Treatment efficacy was evaluated by phototrichogram and digital photograph analysis on 6 repeated treatments for 3 months. Results: Phototrichogram of GFC including FGF9 and normal saline treated side on scalp showed 27.1/cm2 and 5.4/cm2 increase in hair density and 2.7 μm and -0.6μm change in hair diameter, respectively. These results were statistically significant except hair diameter in normal saline side. In density and diameter, GFC including FGF9 was significantly more effective than in normal saline. GFC including FGF9 treatment was compared with ordinary GFC which is a GFC without FGF9, and was more effective in hair density compared to ordinary GFC. Conclusion: GFC including FGF9 treatment with microneedling was effective and safe and it seems to be more effective than ordinary GFC. More research is necessary to determine the most effective factor with various factors of GFC.