Liu Gongyan, Bai Liya, Li Shu, Liu Hongli, Zhu Yanli, Sun Haitao, Gao Shuxia, Jiang Wenxue, Li Fuchang
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.
Animal Husbandry and Veterinary Institute, Shandong Academy of Agricultural Sciences, Shandong Key Laboratory of Animal Disease Control and Breeding, Jinan, 251000, PR China.
Tissue Cell. 2020 Aug;65:101348. doi: 10.1016/j.tice.2020.101348. Epub 2020 Feb 27.
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 day and the 7 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.
真皮乳头细胞（DPCs）是毛囊的关键真皮成分，直接调节毛囊的发育、生长和再生。成功分离培养獭兔的DPCs可为体外研究毛囊发育机制提供良好模型。从30日龄獭兔采集皮肤样本，通过Dispase II和胶原酶D联合使用进行分离，对DPCs进行分离、培养和纯化。观察体外DPCs的形态并绘制生长曲线，DPCs数量在第4天至第7天呈对数模型逐步增加。免疫化学和免疫荧光结果显示细胞中α平滑肌肌动蛋白（α-SMA）和多功能蛋白呈阳性。使用MTT在24小时、48小时、72小时、96小时、120小时和144小时观察第3代（P3）、P6、P9和P12代DPCs的生长特性。P12的细胞密度低于P3（P<0.05）；流式细胞术分析显示，P3处于静止期/第一间隙（G0/G1）期的DPCs高于P12（P<0.05），P3处于第二间隙/有丝分裂（G2/M）期的低于P12（P<0.05）。然而，P12的DPCs呈三角形或短梭形，保留其独特的聚集生长特性。该结果表明，獭兔P12的DPCs特性仍适合体外功能研究。总之，我们成功建立了獭兔DPCs的培养条件，并为研究毛囊发育的分子机制提供了材料。
