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

Gongyan Liu; Liya Bai; Shu Li; Hongli Liu; Yanli Zhu; Haitao Sun; Shuxia Gao; Wenxue Jiang; Fuchang Li

doi:10.1016/j.tice.2020.101348

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

Tissue Cell. 2020 Aug:65:101348. doi: 10.1016/j.tice.2020.101348. Epub 2020 Feb 27.

Authors

Gongyan Liu¹, Liya Bai², Shu Li³, Hongli Liu³, Yanli Zhu³, Haitao Sun², Shuxia Gao², Wenxue Jiang², Fuchang Li⁴

Affiliations

¹ Animal Husbandry and Veterinary Institute, Shandong Academy of Agricultural Sciences, Shandong Key Laboratory of Animal Disease Control and Breeding, Jinan, 251000, PR China; College of Animal Science and Technology, Shandong Agricultural University, Shandong Provincial Key Laboratory of Animal Biotechnology and Disease Control and Prevention, Tai'an, 271018, PR China.
² Animal Husbandry and Veterinary Institute, Shandong Academy of Agricultural Sciences, Shandong Key Laboratory of Animal Disease Control and Breeding, Jinan, 251000, PR China.
³ College of Animal Science and Technology, Shandong Agricultural University, Shandong Provincial Key Laboratory of Animal Biotechnology and Disease Control and Prevention, Tai'an, 271018, PR China.
⁴ College of Animal Science and Technology, Shandong Agricultural University, Shandong Provincial Key Laboratory of Animal Biotechnology and Disease Control and Prevention, Tai'an, 271018, PR China. Electronic address: chlf@sdau.edu.cn.

PMID: 32746997
DOI: 10.1016/j.tice.2020.101348

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 4^th day and the 7^th 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.

Keywords: Cell isolation; Dermal papilla cell; Growth characteristics; Rex rabbit.

MeSH terms

Animals
Biomarkers / metabolism
Cell Cycle
Cell Proliferation
Cell Separation*
Cell Shape
Cells, Cultured
Cryopreservation
Dermis / cytology*
Hair Follicle / cytology
Rabbits
Staining and Labeling
Versicans / metabolism

Substances

Biomarkers
Versicans