Subscribe to RSS
DOI: 10.1055/s-0035-1571191
Insulin Cannot Induce Adipogenic Differentiation in Primary Cardiac Cultures
Publication History
23 September 2015
30 November 2015
Publication Date:
14 January 2016 (online)

Abstract
Cardiac tissue contains a heterogeneous population of cardiomyocytes and nonmyocyte population especially fibroblasts. Fibroblast differentiation into adipogenic lineage is important for fat accumulation around the heart which is important in cardiac pathology. The differentiation in fibroblast has been observed both spontaneously and due to increased insulin stimulation. The present study aims to observe the effect of insulin in adipogenic differentiation of cardiac cells present in primary murine cardiomyocyte cultures.
Oil Red O (ORO) staining has been used for observing the lipid accumulations formed due to adipogenic differentiation in murine cardiomyocyte cultures. The accumulated lipids were quantified by ORO assay and normalized using protein estimation. The lipid accumulation in cardiac cultures did not increase in presence of insulin. However, addition of other growth factors like insulin-like growth factor 1 and epidermal growth factor promoted adipogenic differentiation even in the presence of insulin and other inhibitory molecules such as vitamins. Lipid accumulation also increased in cells grown in media without insulin after an initial exposure to insulin-containing growth media.
The current study adds to the existing knowledge that the insulin by itself cannot induce adipogenic induction in the cardiac cultures. The data have significance in the understanding of cardiovascular health especially in diabetic patients.
-
References
- 1 Parameswaran S, Kumar S, Verma RS, Sharma RK. Cardiomyocyte culture - an update on the in vitro cardiovascular model and future challenges. Can J Physiol Pharmacol 2013; 91 (12) 985-998
- 2 Louch WE, Sheehan KA, Wolska BM. Methods in cardiomyocyte isolation, culture, and gene transfer. J Mol Cell Cardiol 2011; 51 (3) 288-298
- 3 Deb A, Ubil E. Cardiac fibroblast in development and wound healing. J Mol Cell Cardiol 2014; 70: 47-55
- 4 Lajiness JD, Conway SJ. Origin, development, and differentiation of cardiac fibroblasts. J Mol Cell Cardiol 2014; 70: 2-8
- 5 Souders CA, Bowers SLK, Baudino TA. Cardiac fibroblast: the renaissance cell. Circ Res 2009; 105 (12) 1164-1176
- 6 Sacks HS, Fain JN. Human epicardial adipose tissue: a review. Am Heart J 2007; 153 (6) 907-917
- 7 Yamaguchi Y, Cavallero S, Patterson M , et al. Adipogenesis and epicardial adipose tissue: a novel fate of the epicardium induced by mesenchymal transformation and PPARγ activation. Proc Natl Acad Sci U S A 2015; 112 (7) 2070-2075
- 8 d'Amati G, di Gioia CR, Giordano C, Gallo P. Myocyte transdifferentiation: a possible pathogenetic mechanism for arrhythmogenic right ventricular cardiomyopathy. Arch Pathol Lab Med 2000; 124: 287-290
- 9 Pantanowitz L. Fat infiltration in the heart. Heart 2001; 85 (3) 253
- 10 Lombardi R, Chen SN, Ruggiero A , et al. Abstract 20201: Fibro-adipocyte progenitors are a cell source of adipocytes in arrhythmogenic cardiomyopathy. Circulation 2014; 130: A20201
- 11 Zhang Y, Li T-S, Lee S-T , et al. Dedifferentiation and proliferation of mammalian cardiomyocytes. PLoS ONE 2010; 5 (9) e12559
- 12 Chang Y, Li H, Guo Z. Mesenchymal stem cell-like properties in fibroblasts. Cell Physiol Biochem 2014; 34 (3) 703-714
- 13 Driesen RB, Nagaraju CK, Abi-Char J , et al. Reversible and irreversible differentiation of cardiac fibroblasts. Cardiovasc Res 2014; 101 (3) 411-422
- 14 Cieslik KA, Trial J, Carlson S, Taffet GE, Entman ML. Aberrant differentiation of fibroblast progenitors contributes to fibrosis in the aged murine heart: role of elevated circulating insulin levels. FASEB J 2013; 27 (4) 1761-1771
- 15 Bernal A, Gálvez BG. The potential of stem cells in the treatment of cardiovascular diseases. Stem Cell Rev 2013; 9 (6) 814-832
- 16 Sreejit P, Verma RS. Natural ECM as biomaterial for scaffold based cardiac regeneration using adult bone marrow derived stem cells. Stem Cell Rev 2013; 9 (2) 158-171
- 17 Loo ZX, Kunasekaran W, Govindasamy V, Musa S, Abu Kasim NH. Comparative analysis of cardiovascular development related genes in stem cells isolated from deciduous pulp and adipose tissue. ScientificWorldJournal 2014; 2014: 186508
- 18 Vidyasekar P, Shyamsunder P, Santhakumar R, Arun R, Verma RS. A simplified protocol for the isolation and culture of cardiomyocytes and progenitor cells from neonatal mouse ventricles. Eur J Cell Biol 2015; 94 (10) 444-452
- 19 Parameswaran S, Santhakumar R, Vidyasekar P, Verma RS. Enrichment of cardiomyocytes in primary cultures of murine neonatal hearts. In: Skuse GR, Ferran MC, eds. Cardiomyocytes: Methods and Protocols. New York, NY: Springer; 2015: 17-25
- 20 Sreejit P, Kumar S, Verma RS. An improved protocol for primary culture of cardiomyocyte from neonatal mice. In Vitro Cell Dev Biol Anim 2008; 44 (3–4) 45-50
- 21 White SM, Constantin PE, Claycomb WC. Cardiac physiology at the cellular level: use of cultured HL-1 cardiomyocytes for studies of cardiac muscle cell structure and function. Am J Physiol Heart Circ Physiol 2004; 286 (3) H823-H829
- 22 Ramírez-Zacarías JL, Castro-Muñozledo F, Kuri-Harcuch W. Quantitation of adipose conversion and triglycerides by staining intracytoplasmic lipids with Oil red O. Histochemistry 1992; 97 (6) 493-497
- 23 Parameswaran S, Verma RS. Scanning electron microscopy preparation protocol for differentiated stem cells. Anal Biochem 2011; 416 (2) 186-190
- 24 Klemm DJ, Leitner JW, Watson P , et al. Insulin-induced adipocyte differentiation. Activation of CREB rescues adipogenesis from the arrest caused by inhibition of prenylation. J Biol Chem 2001; 276 (30) 28430-28435
- 25 Reusch JEB, Colton LA, Klemm DJ. CREB activation induces adipogenesis in 3T3-L1 cells. Mol Cell Biol 2000; 20 (3) 1008-1020
- 26 El-Jack AK, Hamm JK, Pilch PF, Farmer SR. Reconstitution of insulin-sensitive glucose transport in fibroblasts requires expression of both PPARgamma and C/EBPalpha. J Biol Chem 1999; 274 (12) 7946-7951
- 27 Hainque B, Guerre-Millo M, Hainault I, Moustaid N, Wardzala LJ, Lavau M. Long term regulation of glucose transporters by insulin in mature 3T3-F442A adipose cells. Differential effects on two glucose transporter subtypes. J Biol Chem 1990; 265 (14) 7982-7986
- 28 Aguiari P, Leo S, Zavan B , et al. High glucose induces adipogenic differentiation of muscle-derived stem cells. Proc Natl Acad Sci U S A 2008; 105 (4) 1226-1231
- 29 Jones DD, Ramsay TG, Hausman GJ, Martin RJ. Norepinephrine inhibits rat pre-adipocyte proliferation. Int J Obes Relat Metab Disord 1992; 16 (5) 349-354
- 30 Yamashita H, Sato N, Kizaki T , et al. Norepinephrine stimulates the expression of fibroblast growth factor 2 in rat brown adipocyte primary culture. Cell Growth Differ 1995; 6 (11) 1457-1462
- 31 Kakudo N, Shimotsuma A, Kusumoto K. Fibroblast growth factor-2 stimulates adipogenic differentiation of human adipose-derived stem cells. Biochem Biophys Res Commun 2007; 359 (2) 239-244
- 32 Garten A, Schuster S, Kiess W. The insulin-like growth factors in adipogenesis and obesity. Endocrinol Metab Clin North Am 2012; 41 (2) 283-295 , v–vi
- 33 Harrington M, Pond-Tor S, Boney CM. Role of epidermal growth factor and ErbB2 receptors in 3T3-L1 adipogenesis. Obesity (Silver Spring) 2007; 15 (3) 563-571
- 34 Dasgupta S, Bhattacharya S, Biswas A , et al. NF-kappaB mediates lipid-induced fetuin-A expression in hepatocytes that impairs adipocyte function effecting insulin resistance. Biochem J 2010; 429 (3) 451-462
- 35 Wlazlo N, van Greevenbroek MMJ, Ferreira I , et al. Iron metabolism is associated with adipocyte insulin resistance and plasma adiponectin: the Cohort on Diabetes and Atherosclerosis Maastricht (CODAM) study. Diabetes Care 2013; 36 (2) 309-315
- 36 Rahman F, Al Frouh F, Bordignon B , et al. Ascorbic acid is a dose-dependent inhibitor of adipocyte differentiation, probably by reducing cAMP pool. Front Cell Dev Biol 2014; 2: 29
- 37 Berry DC, DeSantis D, Soltanian H, Croniger CM, Noy N. Retinoic acid upregulates preadipocyte genes to block adipogenesis and suppress diet-induced obesity. Diabetes 2012; 61 (5) 1112-1121
- 38 Madsen L, Petersen RK, Kristiansen K. Regulation of adipocyte differentiation and function by polyunsaturated fatty acids. Biochim Biophys Acta 2005; 1740 (2) 266-286