Background: Growth Factor Cocktail (GFC), which contains growth factors and cytokine, has been gradually gaining attention as a new treatment for hair loss. Several studies reported the effectiveness of GFC on hair regrowth via microneedles; however, studies regarding formulations with fibroblast growth factor 5 short (FGF5s) and nicotinamide mononucleotide (NMN) have not been conducted. Objectives: This study aimed to investigate the effect of treatment with microneedle technology by adding FGF5s and NMN to GFC in patients with androgenetic alopecia (AGA). Methods: In this study, AGA patients were treated six times for a total of 3 months at 2-week intervals from the first visit. The scalp was divided into two sections at the time of treatment; the right side was treated with GFC with FGF5s and NMN, and the left were treated with normal saline, with microneedles to a depth of 0.8 mm. A total of 20 patients (11 males and 9 females) were included in the study. Clinical photographs and phototrichogram were performed before and 2 weeks after treatment to determine the final treatment effect. Results: The phototrichogram images of the scalp treated with GFC for 3 months showed increased hair density and diameter. In contrast, there was no change in hair density and diameter in the scalp treated with normal saline. Conclusion: GFC with FGF5s and NMN with microneedling is an effective and safe treatment of androgenetic alopecia. Future studies in controlled clinical settings are warranted.