Premium
Characterization of a novel murine preadipocyte line, AP‐18, isolated from subcutaneous tissue: Analysis of adipocyte‐related gene expressions
Author(s) -
Chen Chen,
Takahashi Kazuma,
Yoshida Ayumi,
Takizawa Yumiko,
Lee Yan,
Nakui Masaki,
Doi Hideyuki,
Takebayashi Yuji,
Fukumoto Manabu,
Yamada Testuya,
Katagiri Hideki,
Oka Yoshitomo,
Satoh Jo
Publication year - 2010
Publication title -
cell biology international
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.932
H-Index - 77
eISSN - 1095-8355
pISSN - 1065-6995
DOI - 10.1042/cbi20090063
Subject(s) - adipocyte , glut4 , adipose tissue , 3t3 l1 , cell culture , adiponectin , medicine , endocrinology , biology , cellular differentiation , lipid droplet , white adipose tissue , leptin , microbiology and biotechnology , chemistry , insulin , glucose transporter , insulin resistance , gene , biochemistry , genetics , obesity
Adipocyte lines are a useful tool for adipocyte research. Recently, a new preadipocyte line designated AP‐18 was established from subcutaneous tissue of the C3H/He mouse. In this study, we further characterized AP‐18 cells. Adipocyte differentiation was assessed by accumulation of fat droplets stained by Oil Red O. The expression of the preadipocyte‐ or adipocyte‐specific genes and adipocytokine genes was analysed qualitatively by RT‐PCR and quantitatively by real‐time PCR in comparison with the LM cell, a murine fibroblast line, and the 3T3‐L1 cell, respectively. AP‐18 cells were fibroblastoid in maintenance culture. After the confluence, fat droplets were accumulated in 50–60% of the cells cultured in the medium alone and in 70–90% of the cells cultured with insulin within 2 to 3 weeks. The fat accumulation was not promoted by the addition of dexamethazone, IBMX (3‐isobutyl‐1‐methylxanthine) or troglitazone in combination with insulin, which were obligatory for differentiation of the 3T3‐L1 cell, a murine preadipocyte line. Throughout the differentiation, AP‐18 cells expressed Pref‐1, LPL, C/EBPβ, C/EBPδ, RXRα, C/EBPα, PPARγ, RXRγ, aP2, GLUT4, SCD1, UCP2, UCP3, TNFα, resistin, leptin, adiponectin and PAI‐1 genes, but not the UCP1 gene, indicating that the cell is derived from WAT (white adipose tissue). The time course of these gene expressions was similar to that of 3T3‐L1 cells, although the expressions were slower and lower in AP‐18 cells. These data indicate that AP‐18 cells are preadipocytes originated from WAT and differentiate into adipocytes under more physiological conditions than 3T3‐L1 cells. AP‐18 may be useful in adipocyte research.