Development of hair follicle spheroids with growth factor transfection and various cell combinations

Abstract

As the number of hair loss patients increases, research conducted on hair follicles and hair has recently become more active. However, most current hair follicle research models are limited in their ability to reproduce several key functions of the hair follicle microenvironment. To solve this problem, hair growth genes were injected into hair follicle cells, and long-term culture was performed for up to 90 days. DPCs (dermal papilla cells), KCs (keratinocytes), HUVECs (human umbilical vein endothelial cells) or HDMECs (human dermal microvascular endothelial cells), and FBs (fibroblasts) injected with hair follicle growth factors LEF1 and Wnt1 were mixed to produce spheroids, and the growth factor expression and growth behavior according to the type of cell and the presence or absence of transfection according to the culture period were analyzed using SEM, H&E staining, and IF imaging. As the culture period increased, the size of the spheroids increased, and the number decreased due to the association between spheroids; long fibers that appeared to be hair expression were observed. We also compared the differences between HUVECs and HDMECs, which have different phenotypes of vascular cells. In spheroids that did not contain FBs, the expression of HUVECs was more promoted, whereas in spheroids that contained FBs, the expression of HDMECs tended to be more promoted. These results are expected to contribute to dermatology and new drug development for the prevention of intractable skin diseases and hair loss using skin models with a microbiome environment in the future.