Suppression of adipogenic differentiation by muscle cell‐induced decrease in genes related to lipogenesis in muscle and fat co‐culture system

Abstract

Intercellular signalling communication between adipose and muscle tissue has been investigated. To test the effect of muscle cells on adipogenic gene expression, we utilised an in vitro co-culture system, in which fat (3T3-L1) and muscle (L-6) cells were physically separated but chemically exposed each other via insert with 0.4 µm porous membrane. When 3T3-L1 and L-6 cells reached at 80 and 40% confluence, respectively in separate wells, L-6 cells grown in insert were transferred onto 6-well plates where 3T3-L1 cells were being grown. When both cells were fully differentiated in co-culture plates, morphology of 3T3-L1 was examined by staining with Oil-red-O. Activity of glycerol-3-phosphate dehydrogenase (GPDH) and adipogenic gene expression including lipoprotein lipase (LPL), adipsin, GPDH, peroxisome proliferator-activated receptor-γ (PPARγ) and CCAAT/enhancer binding protein (C/EBPα) were analysed. The presence of muscle cells during preadipocyte differentiation inhibited (P < 0.05) lipogenesis by suppressing lipogenic gene expression including LPL, adipsin and GPDH. Furthermore, GPDH activity was also decreased (P < 0.05) in 3T3-L1 cells by the presence of L-6 cells. These results suggest that presence of muscle cells suppresses adipogenic differentiation by inhibiting the adipogenic gene expression and GPDH activity in the muscle and fat cell co-culture system.

AMPKα, C/EBPβ, CPT1β, GPR43, PPARγ, and SCD Gene Expression in Single- and Co-cultured Bovine Satellite Cells and Intramuscular Preadipocytes Treated with Palmitic, Stearic, Oleic, and Linoleic Acid

“…Most studies of the regulation of adipogenesis have been conducted in cultured primary preadipocyte cultures or 3T3-L1 secondary preadipocyte cultures. A few studies have investigated the differentiation of bovine adipocytes and myoblasts in single cultures ( Chung et al, 2006a ; Ouellette et al, 2009 ), and we are aware of three studies describing the interaction between adipocytes and myoblastic cells in a co-culture system ( Hausman et al, 2005 ; Choi et al, 2013a ; Park et al, 2013 ), two of which originated from our laboratory ( Choi et al, 2013a ; Park et al, 2013 ). We have used this set of genetic markers of adipose tissue differentiation ( C/EBPβ , PPARγ ) and lipid metabolism ( AMPKα , CPT1β , GPR43 , SCD ) in studies of the effects of fatty acids on bovine preadipocytes ( Chung et al, 2006a ) and mature adipose tissues ( Smith et al, 2012 ; Choi et al, 2013a ).…”

Adipocytes suppress differentiation of muscle cells in a co‐culture system

“…Results from co-culture systems have shown that adipocytes induce insulin resistance (Dietze et al, 2002), and inhibit differentiation (Takegahara, Yamanouchi, Nakamura, Nakano, & Nishihara, 2014) and atrophy (Pellegrinelli et al, 2015) of muscle cells. Furthermore, adipogenesis of mesenchymal adipocyte progenitors or 3T3-L1 cells is inhibited by the presence of muscle satellite cells or L6 muscle cells (Park, Baek, & Choi, 2013;Uezumi et al, 2010). However, it is still unclear how myogenic and adipogenic cells communicate with each other in the extracellular niche within skeletal muscle.…”

The Evolution of Complex Muscle Cell In Vitro Models to Study Pathomechanisms and Drug Development of Neuromuscular Disease

“…In a mouse model for DMD ( mdx mouse), they seem to provide the main source of increased fibrogenic or adipose tissue in pathological muscle [ 123 ]. Interestingly, a co-culture of fat (3T3-L1) and muscle (L-6) in physical separation but with chemically reciprocal exposure resulted in suppressed lipogenic gene expression and reduced GPDH-activity, suggesting an inhibitory influence of skeletal muscle on adipogenic differentiation [ 124 ]. A co-culture of human fat and skeletal muscle cells, obtained from different muscle cell and fat cell donors, showed induction of insulin resistance in human muscle cells through the release of fat cell factors [ 125 , 126 ], underlining the alternating influence of muscle on fat and vice versa.…”