title: Co-culture of bovine muscle satellite cells with preadipocytes increases PPARγ and C/EBPβ gene expression in differentiated myoblasts and increases GPR43 gene expression in adipocytes.
abstract: We hypothesized that preadipocyte differentiation would be depressed by differentiating myoblasts, whereas preadipocytes would promote adipogenic gene expression in myoblasts in a co-culture system. We also determined the effects of arginine, a biological precursor of nitric oxide, and/or trans-10, cis-12 conjugated linoleic acid (CLA) on adipogenic gene expression during differentiation of bovine preadipocytes and myoblasts. Bovine semimembranosus satellite cells (BSC) and subcutaneous preadipocytes were isolated from crossbred steers and cultured with 10% fetal bovine serum (FBS)/Dulbecco's modified Eagle medium (DMEM) and 1% antibiotics during the 3-day proliferation period. After proliferation, BSC and preadipocytes were treated for 3 days with 3% horse serum/DMEM and 5% FBS/DMEM with antibiotics, respectively. Media also contained 100 μM oleic acid, 10 μg/ml insulin, 1 μg/ml pioglitazone and 1 μg/ml dexamethasone. Subsequently, the differentiating myoblasts and adipocytes were cultured in their respective media with 5 mM arginine and/or 40 μM trans-10, cis-12 CLA for 4 days. Finally, myoblasts and adipocytes were single- or co-cultured for 2 h singly or in combination. Arginine stimulated SCD gene expression, whereas CLA depressed SCD gene expression in adipocytes and myoblasts (P=.002). Co-culture of adipocytes and myoblasts elicited an increase in C/EBPβ and PPARγ gene expression in differentiated myoblasts (P≤.01) and an increase in GPR43 gene expression in adipocytes (P=.01). Expression of AMPKα and CPT1ß was unaffected by co-culture, although SCD gene expression tended (P=.12) to be depressed by co-culture. These experiments demonstrated that co-culture of adipocytes with myoblasts increased adipogenic gene expression in the myoblastic cells.
authors: Department of Animal Science, Texas Agricultural Experiment Station, Texas A&M University, College Station, TX 77843, USA.
authors: Seong H
authors: SH
authors: Choi
authors: Ki Y
authors: KY
authors: Chung
authors: Bradley J
authors: BJ
authors: Johnson
authors: Gwang W
authors: GW
authors: Go
authors: Kyoung H
authors: KH
authors: Kim
authors: Chang W
authors: CW
authors: Choi
authors: Stephen B
authors: SB
authors: Smith
mesh_terms: D055372:AMP-Activated Protein Kinases / Q000235:genetics / Q000378:metabolism; D017667:Adipocytes / Q000166:cytology* / Q000378:metabolism; D000818:Animals; D022782:CCAAT-Enhancer-Binding Protein-beta / Q000235:genetics / Q000378:metabolism*; D002417:Cattle; D002454:Cell Differentiation; D049109:Cell Proliferation; D002478:Cells, Cultured; D018920:Coculture Techniques; D003907:Dexamethasone / Q000032:analysis / Q000378:metabolism; D015870:Gene Expression; D005786:Gene Expression Regulation; D007328:Insulin / Q000032:analysis / Q000378:metabolism; D044243:Linoleic Acids, Conjugated / Q000378:metabolism; D008297:Male; D019301:Oleic Acid / Q000032:analysis / Q000378:metabolism; D047495:PPAR gamma / Q000235:genetics / Q000378:metabolism*; D000077205:Pioglitazone; D043562:Receptors, G-Protein-Coupled / Q000235:genetics / Q000378:metabolism*; D032496:Satellite Cells, Skeletal Muscle / Q000166:cytology* / Q000378:metabolism; D045162:Thiazolidinediones / Q000032:analysis / Q000378:metabolism
chemical_list: D022782:CCAAT-Enhancer-Binding Protein-beta; D007328:Insulin; D044243:Linoleic Acids, Conjugated; D047495:PPAR gamma; D043562:Receptors, G-Protein-Coupled; D045162:Thiazolidinediones; C046938:9,11-linoleic acid; D019301:Oleic Acid; D003907:Dexamethasone; D055372:AMP-Activated Protein Kinases; D000077205:Pioglitazone