Characterization of a novel murine preadipocyte line, AP-18, isolated from subcutaneous tissue: analysis of adipocyte-related gene expressions.
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/EBP beta, C/EBP delta, RXR alpha, C/EBP alpha, PPAR gamma, RXR gamma, aP2, GLUT4, SCD1, UCP2, UCP3, TNFalpha, 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.
Chen
Chen
C
Division of Molecular Metabolism and Diabetes, Tohoku University, Sendai, Japan.
Takahashi
Kazuma
K
Yoshida
Ayumi
A
Takizawa
Yumiko
Y
Lee
Yan
Y
Nakui
Masaki
M
Doi
Hideyuki
H
Takebayashi
Yuji
Y
Fukumoto
Manabu
M
Yamada
Testuya
T
Katagiri
Hideki
H
Oka
Yoshitomo
Y
Satoh
Jo
J
D041721:3T3-L1 Cells; D017667:Adipocytes / Q000166:cytology / Q000378:metabolism*; D052242:Adiponectin / Q000235:genetics / Q000378:metabolism; D052436:Adipose Tissue, White / Q000378:metabolism; D000818:Animals; D002135:Calcium-Binding Proteins; D002454:Cell Differentiation; D002460:Cell Line; D005786:Gene Expression Regulation; D051275:Glucose Transporter Type 4 / Q000235:genetics / Q000378:metabolism; D036341:Intercellular Signaling Peptides and Proteins / Q000235:genetics / Q000378:metabolism; D020738:Leptin / Q000235:genetics / Q000378:metabolism; D058867:Lipoprotein Lipase Activators; D051379:Mice; D008809:Mice, Inbred C3H; D047495:PPAR gamma / Q000235:genetics / Q000378:metabolism; D010455:Peptides / Q000235:genetics / Q000378:metabolism; D017395:Plasminogen Activator Inhibitor 1 / Q000235:genetics / Q000378:metabolism; D052243:Resistin / Q000235:genetics / Q000378:metabolism; D047490:Retinoid X Receptor alpha / Q000235:genetics / Q000378:metabolism; D013234:Stem Cells / Q000378:metabolism*; D040521:Subcutaneous Tissue / Q000033:anatomy & histology*; D013997:Time Factors
D052242:Adiponectin; D002135:Calcium-Binding Proteins; C512925:Dlk1 protein, mouse; D051275:Glucose Transporter Type 4; D036341:Intercellular Signaling Peptides and Proteins; D020738:Leptin; D058867:Lipoprotein Lipase Activators; D047495:PPAR gamma; D010455:Peptides; D017395:Plasminogen Activator Inhibitor 1; D052243:Resistin; D047490:Retinoid X Receptor alpha