The proliferation and differentiation characteristics of co-cultured porcine preadipocytes and muscle satellite cells in vitro

Abstract

To explore the proliferation and differentiation characteristics of co-cultured porcine preadipocytes and muscle satellite cells, preadipocytes and muscle satellite cells were isolated from the healthy nascent landrace. Oil Red O stain and desmin immunohistochemistry were used to identify the two solo-cultured cells. Methyl thiazolyl tetrazolium (MTT) colorimetric assay was used to detect the proliferation characteristic of co-cultured cells, and the expression level of differentiation marker genes lipoprotein lipase (LPL), peroxisome proliferator-activated receptors (PPARγ), myogenic factor 5 (Myf5), myogenin (MyoG) were analyzed with reverse transcription PCR (RT-PCR) and western blot. The success of co-culture system was proved. In the co-cultured cells, slight lipid droplets were observed and appeared more slowly. The polykaryocytes fused into myotubes in co-cultured cells were less and relatively slow than that in solo myocytes. After fusion, the proliferation rate of co-cultured cells was higher than that in the solo-cultured muscle satellite cells (P < 0.01), and the duration were also longer. On day 5 and 10, the expression of the marker genes in earlier stage of cell differentiation (LPL and Myf5) were lower than those in the solo-cultured cells (P < 0.01) (except LPL gene at day 5). Moreover, the expression of intermediate and advanced stages' maker genes (PPARγ2 and MyoG) were hardly detectable at day 5, but increased significantly on day 10 (P < 0.01). These results confirm that the co-culture system could facilitate the cells' growth and proliferation, meanwhile, inhibited the cell differentiation.

Muscle-specific downregulation of GR levels inhibits adipogenesis in porcine intramuscular adipocyte tissue

“…Most studies of muscle-adipose crosstalk were focused on the effects of adipose on the muscle, only a few studies reported that myocytes suppressed differentiation of adipocytes 43, 44 , and degenerating myofibres could affected the cellular morphology of 3T3-L1 preadipocytes 45 . However, the mechanism was still unknown.…”

Molecular and Cellular Response of Co-cultured Cells toward Cobalt Chloride (CoCl2)-Induced Hypoxia

“…34 In addition, the differentiation of porcine preadipocytes was repressed in the co-culture system and the expression of marker genes in the previous phase of adipogenic differentiation was lesser than that in the monocultured cells. 35…”

Isolation and osteogenic differentiation of skeletal muscle-derived stem cells for bone tissue engineering

“…Skeletal muscle has been extensively investigated as a potential source for isolation of several stem cells over the past few years (6). Satellite cells located beneath the basal lamina of mature skeletal muscle fibers, often referred to as 'muscle stem cells', have long been considered monopotential stem cells capable of only giving rise to cells of the myogenic lineage (7). MDSCs, which may represent a predecessor of the satellite cell, are considered to be distinct in that they may not be restricted to the myogenic lineage or mesenchymal tissues and are able to differentiate into multiple lineages (8).…”