Abstract
Androgenetic alopecia (AGA) is the most common type of hair loss, and is mainly caused by the biological effects of testosterone on dermal papilla cells (DPCs). In vitro culturing of DPCs might be a useful tool for the screening of target molecule of AGA. However, primary DPCs cannot continuously proliferate owing to cellular senescence and cell culture stress. In this study, we introduced mutant cyclin-dependent kinase 4 (CDK4), Cyclin D1, and telomerase reverse transcriptase (TERT) into DPCs. We confirmed protein expression of CDK4 and Cyclin D1, and enzymatic activity of TERT. Furthermore, we found the established cell line was free from cellular senescence. We also introduced the androgen receptor gene using a recombinant retrovirus, to compensate the transcriptional suppressed endogenous androgen receptor in the process of cell proliferation. Furthermore, we detected the efficient nuclear translocation of androgen receptor into the nucleus after the treatment of dihydrotestosterone, indicating the functionality of our introduced receptor. Our established cell line is a useful tool to identify the downstream signaling pathway, which activated by the testosterone.
Highlights
Hair plays an important role in protecting the skin from mechanical damage
We named the cells transfected with R24C mutant cyclin-dependent kinase 4 (CDK4), Cyclin D1, and telomerase reverse transcriptase (TERT) as K4DT cells, in reference to the modified genes (Katayama et al, 2019)
We named the recombinant cells with R24C mutant CDK4 and Cyclin D1 as K4D cells
Summary
Hair plays an important role in protecting the skin from mechanical damage. The hair follicle regulates hair growth through complex interactions between hormones, neuropeptides, and immune cells. The hair follicle consists of multiple cell types, including dermal papilla cells (DPCs), matrix cells, and melanocytes (Driskell et al, 2011). The hair follicle remains dormant while new hair growth begins, eventually causing hair loss or “shedding” of the old hair. This hair growth and replacement process is known as the hair cycle (Davis, 1962). Hair growth and proliferation is mainly due to the hair matrix cell proliferation and Abbreviations: AGA, Androgenetic alopecia; DPCs, dermal papilla cells; CDK, 4cyclin-dependent kinase 4; TERT, telomerase reverse transcriptase; DHT, dihydrotestosterone; TGFβ, transforming growth factor β; Dkk, Dickkopf-related protein 1; DAPI, 4 ,6-diamidino-2-phenylindole; SA-β-Gal, senescence-associated β-galactosidase; AP, alkaline phosphatase
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