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

BioMed Research International
Man-Feng Zhang Ge-Shi

Abstract

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.

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Citations

Mar 12, 2020·Clinical, Cosmetic and Investigational Dermatology·James Nicoll, Benjamin M Buehrer
Sep 15, 2020·The Journal of Veterinary Medical Science·Akira MatsudaYoshiki Itoh

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Methods Mentioned

BETA
FACS
protein assay
FCS
flow cytometry
transmission electron microscopy
gene knockdown

Software Mentioned

ModFit LT
Proteome Discoverer
Mascot

Related Concepts

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