Differentiation Model Establishment and Differentiation-Related Protein Screening in Primary Cultured Human Sebocytes

• Main Authors: Man-Feng Zhang, Xiao-Lin Cai, Kai-Peng Jing, Xiao-Xue Pi, Pei-Yu Liao, Shi-Jie Li, Wen-Li, Chuan-Chuan Cai, Juan-Hua Quan, Yi-Ming Fan, Ge-Shi
• Format: Article
• Language: English
• Published: Hindawi Limited 2018-01-01
• Series: BioMed Research International
• Online Access: http://dx.doi.org/10.1155/2018/7174561

Sebocyte differentiation is a continuous process, but its potential molecular mechanism remains unclear. We aimed to establish a novel sebocyte differentiation model using human primary sebocytes and to identify the expression profiles of differentiation-associated proteins. Primary human sebocytes were cultured on Sebomed medium supplemented with 2% serum for 7 days. Flow cytometry showed that S phase cells were decreased time-dependently, while G1 and subG1 (apoptosis) phase cells increased under serum starvation. Transmission electron microscopy and Oil Red O staining revealed a gradual increase of intracellular lipid accumulation. Expression of proliferation marker was diminished, while expression of differentiation, apoptosis, and lipogenic markers elevated gradually during 7-day culture. iTRAQ analysis identified 3582 expressed proteins in this differentiation model. Compared with day 0, number of differentially expressed proteins was 132, 54, 321, and 96 at days 1, 3, 5, and 7, respectively. Two overexpressed proteins (S100 calcium binding protein P and ferredoxin reductase) and 2 downexpressed proteins (adenosine deaminase and keratin 10) were further confirmed by Western blot and immunohistochemistry.