The proliferation and differentiation of primary pig preadipocytes is suppressed when cultures are incubated at 37°Celsius compared to euthermic conditions in pigs

Abstract

Given similarities in metabolic parameters and cardiovascular physiology, the pig is well positioned as a biomedical model for metabolic disease and obesity in humans. Better understanding molecular mechanisms governing porcine adipocyte hyperplasia may provide insight into the regulation of adipose tissue development that is useful both when considering the pig as a commodity and when extrapolating porcine data to human disease. Primary cultures of pig stromal-vascular cells have served as a useful tool for investigating factors that regulate preadipocyte proliferation and differentiation. However, such cultures have generally been maintained at 37°C in vitro despite euthermia being 39°C in pigs. To address potential concerns about the physiological relevance of culturing primary pig preadipocytes under what would be hypothermic conditions in vivo, the objective of this study was to investigate the effect of culture temperature on the proliferation and differentiation of pig preadipocytes in primary culture. Culturing primary preadipocytes at 37 rather than 39°C decreases their proliferation rates based upon cleavage of the tetrazolium salt, MTT (P < 0.001), reduction of resazurin (P < 0.001), and daily cell counts (P < 0.001). Likewise, culturing primary porcine preadipocytes at 37°C suppressed their adipogenic potential based upon monitoring adipogenesis morphologically, biochemically, and via the expression of mRNA encoding adipogenic marker genes. Collectively, these data indicate the proliferation and differentiation of primary pig preadipocytes is suppressed when cultures are incubated at 37°C compared to normal body temperature of pigs. This may confound investigation of factors that impact adipocyte hyperplasia in the pig.

Dietary fiber sources and non-starch polysaccharide-degrading enzymes modify mucin expression and the immune profile of the swine ileum

“…The culture of swine enteroids, although challenging, has been previously reported in studies showing the presence of all cells of the intestine and their response to bacterial lipopolysaccharide stimulation [ 27 , 53 , 54 ]. The swine enteroids grown in our laboratory showed similar characteristics to those reported in previous studies, although we have defined that culturing at 39°C (the body temperature of pigs) favors the expression of differentiation markers, as has been previously suggested for adipocytes [ 55 ]. There is limited information in the literature about the regulation of mucin in swine.…”

Cell Models and Their Application for Studying Adipogenic Differentiation in Relation to Obesity: A Review

“…In this regard, Shu et al described the role of phloretin, which promotes the differentiation of 3T3-L1 adipocytes, and demonstrated that phloretin enhances the lipid accumulation in a time-dependent manner in porcine primary adipocytes [ 76 ]. In another study, primary cultures of pig SVF cells were differentiated to study the effect of temperature on proliferation and differentiation [ 77 ]. Furthermore, retinol binding protein 4 (RBP-4) was observed to significantly suppress differentiation in porcine preadipocytes by decreasing the activation of insulin signaling pathways [ 78 ].…”

PPARγ agonist through the terminal differentiation phase is essential for adipogenic differentiation of fetal ovine preadipocytes

“…Addition of a PPARγ agonist for 48 h improved differentiation, increasing medium-sized lipid droplet formation (Fig. 2b1) and suggesting that similar to feline [7], porcine [34], and human [31] preadipocytes, ovine preadipocytes have lower sensitivity to insulin than murine cell lines and PPARγ agonist may be a more stringent requirement for adipogenic differentiation in these species.…”

“…They also require higher insulin concentration during differentiation to enhance lipid accumulation and similar to human primary preadipocytes [ 21 , 22 ], PPARγ agonist supplementation is also required for ovine adipogenic differentiation. This work highlights species-specific differences requirements for adipogenic differentiation [ 7 , 14 , 31 , 34 ] and the need to develop standardized methods to investigate comparative adipocyte biology.…”