Isolation, culture and growth characteristics of dermal papilla cells from Rex rabbits

Abstract

Dermal papilla cells (DPCs) is the key dermal component of the hair follicle that directly regulates hair follicle development, growth and regeneration. Successfully isolated and cultured DPCs of Rex rabbit could provide a good model for the study of hair follicle development mechanism in vitro. Skin samples were collected from 30-day old Rex rabbits and separated by combination of Dispase II and Collagenase D, separation, culture, and purification of DPCs. The morphology of DPCs in vitro was observed and the growth curve was drawn, the number of DPCs presented progressive increase in a logarithmic model between the 4th day and the 7th day. The results of immune chemical and immune fluorescence shown that α smooth muscle actin (α-SMA) and versican were positive in cells. Growth character of the passages 3 (P3), P6, P9 and P12 DPCs were observed using MTT at 24 h, 48 h, 72 h, 96 h, 120 h and 144 h. The cell density of P12 was lower than P3 (P < 0.05); the flow cytometric analysis showed that DPCs at resting state/first gap (G0/G1) stage of P3 was higher than P12 (P < 0.05), and second gap/mitosis (G2/M) stage of P3 was lower than P12 (P < 0.05). However, the DPCs of P12 present triangular or short fusiform, retaining their unique aggregative growth characteristics. This results shown that the DPCs properties of P12 from Rex rabbits, still fit functional research in vitro. In conclusion, we successfully established the culturing condition of DPCs from Rex rabbits, and provide a material for studying the molecular mechanism of hair follicle development.