Enzymatic Dissociation and Identification of Hair Follicle Stem Cells for Reversing Miniaturization in Androgenetic Alopecia

<h3>ABSTRACT</h3> <i><b>Introduction:</b></i> The efficacy of hair-follicle-derived stem cells (HFDSCs) in treating androgenetic alopecia (AGA) has been documented in a limited number of studies; however, early results from cell-based therapy offer promising potential for new treatments for hair loss. Unfortunately, the process of dissociating, identifying, and counting these stem cells is complicated and expensive. In this article, we introduce a method for enzymatic dissociation of HFDSCs. We obtained these HFDSCs from transected, partially transected, and healthy hair follicles obtained during hair transplant procedures. The isolated stem cells were then injected into areas of the scalp with miniaturized hair follicles. <i><b>Materials &amp; Methods:</b></i> The study included 20 patients in the test group who received injections of HFDSCs and 20 control patients. Approximately 150 hair follicles (healthy, transected, and/or partially transected) were used for trypsin enzymatic dissociation of stem cells for each patient. The dissociated HFDSCs were mixed in Lactated Ringer’s solution, and the solution from the initial subjects were sent to a laboratory for CD marker and cell count studies to confirm the presence of HFDSCs prior to injection. The CD marker study was conducted using CD200, CD34 (for epithelial stem cells), and CD44 (for mesenchymal stem cells). After confirmation of adequate markers in the initial subjects, the solution was subsequently injected into the scalp areas with miniaturized hair. The subsequent subjects did not have their samples sent to the lab for analysis. <i><b>Results:</b></i> The HFDSC suspension from the initial few subjects showed that 96% of cells were viable. All four samples were positive for CD markers. The test group showed a 19% improvement in hair density (p-value &lt;0.0001) and a 20% improvement in hair caliber (p-value &lt;0.0001) from baseline. Global photographic imaging revealed significant improvement in all test group participants (p-value &lt;0.0001), while the control group experienced significant deterioration in global photographic image grading (p-value &lt;0.0001). <i><b>Conclusion:</b></i> This initial small single-center non-randomized study showed the potential for enzymatic dissociation for the isolation of HFDSCs from hair follicles during hair transplantation and the potential efficacy of injection of this isolate for miniaturized hair in AGA. However, further studies including further study of the isolate with other methodologies and by other laboratories to further analyze and characterize the cell types isolated are still needed. Additionally larger multicenter randomized studies are still needed to further evaluate the safety and efficacy of this treatment modality.