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BIOMIMESYS® Adipose tissue, a relevant in vitro adipocyte 3D model

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BIOMIMESYS® range are hyaluronan based scaffolds developed to overcome the 2D flat culture limitations by recreating an in vivo physiology within the in vitro environment.
BIOMIMESYS® Adipocyte scaffold is made of RGDS-grafted Hyaluronic acid (1.6 MDa), Adipic acid dihydrazide crosslinker and extracellular matrix (ECM) proteins (collagen type I and collagen type VI) to mimic fat tissue-ECM composition.

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BIOMIMESYS® Adipose tissue, a relevant in vitro adipocyte 3D model

  1. 1. BIOMIMESYS®Adipose tissue, a relevant in vitro adipocyte 3D model BIOMIMESYS®Adipose tissue has been tested on 3T3-L1, 3T3-F44 cells and on human cryopreserved preadipocytes (HWP). It makes 3D cell culture easy and provides a robust in vitro and reliable model for metabolism studies such as obesity or diabetes and drug discovery. To know more about BIOMIMESYS®Adipose tissue visit our website: www.biomimesys.com  +33(0) 769 999 137; hello@biomimesys.com BIOMIMESYS® Adipose tissue is ready to use Available in a ready-to-use format (96 well plates) it enables the culture of adipocytes under physiological conditions that are representative of the microenvironment found in adipose tissue(2). Adipocytes cells are simply seeded on top of the hydroscaffold and placed in the incubator. The media can be refreshed easily by pipetting. BIOMIMESYS® Adipose tissue is physiological BIOMIMESYS® range are hyaluronan based hydroscaffolds developed to overcome the 2D flat culture limitations by recreating an in vivo physiology within the in vitro environment. BIOMIMESYS®Adipose tissue hydroscaffold is made of RGDS-grafted Hyaluronic acid (1.6 MDa), Adipic acid dihydrazide crosslinker and extracellular matrix (ECM) proteins (collagen type I and collagen type VI) to mimic fat tissue-ECM composition. In BIOMIMESYS®Adipose tissue PHYSICOCHEMICAL FEATURES Porosity: 70-170 µm Young’s modulus: E = 0.45 ± 0.05kPa Swelling ratio = 60 ± 10g/g Adipocyte maturation BIOMIMESYS®Adipose tissue is compatible with all analytical technologies Being transparent makes it suitable for microscopy (immunofluorescence, brightfield) and use in plate readers (OD, fluorescence). Thanks to its porosity, proteins and nucleic acid can be extracted by directly adding the lysis buffer to the hydrogel. In comparison to in vivo decellularized adipose tissue SEM observation of a human decellularized adipose tissue (from Wang et al., 2013 (1) ) Perilipin immunostaining – HWP a maturation marker : the lipid droplet-associated protein Morphology SEM observation of a BIOMIMESYS®Adipose tissue section, highlighting the collagen chain, (artificially coloured) Biomimetic structure In vivo References: (1) L. Wang et al. Combining decellularized human adipose tissue extracellular matrix and adipose-derived stem cells for adipose tissue engineering, Acta Biomaterialia 2013 (9):8921-8931. (2) G. J. Hausman. Meat Science and Muscle Biology Symposium: The influence of extracellular matrix on intramuscular and extramuscular adipogenesis, Journal of Animal Science 2012 (90):942-949. (3) M. Kurita et al. Influences of Centrifugation on Cells and Tissues in Liposuction Aspirates: Optimized Centrifugation for Lipotransfer and Cell Isolation, Plastic and Reconstructive Surgery 2008 121(3):1033-1041. 50 µm 200 µm Murine Preadipocytes 3T3- L1 at day 7 Human White Preadipocytes (HWP) at day 14 10µm 20µm Human adipocytes clusters from abdomen liposuction (from Kurita et al., 2008 (3) ) In vivo-like organisation of adipose tissue in BIOMIMESYS®Adipose tissue. N=3, two-ways ANOVA statistics with Tukey post-hoc ; * : p<0,05 ; ** : p<0,01 ; *** : p<0,001 *** *** *** ** *** Better lipogenic activity and higher differentiation rate in BIOMIMESYS®Adipose tissue compared to 2D culture. LSCM observation of Z-stack at day 21. Chronic effect on lipogenesis – 3T3-L1 DIFFERENTIATION 48h 72h NUTRITION D0 GROWTH D14D7 D28 ± Retinoic acid (RA) N=3, two-ways ANOVA statistics with Tukey post-hoc ; * : p<0,05 ; ** : p<0,01 ; *** : p<0,001 Drug screening HWP undergo complete maturation using BIOMIMESYS®Adipose tissue. 100µm Perilipin Nucleus Lipids accumulation – 3T3-L1 AdipoRed™ Assay Reagent Adipogenesis - HWP mRNA of a early gene : fatty acid binding protein 4 (FABP4) Adipogenesis - HWP mRNA of a late gene : glucose transporter type 4 (GLUT-4) Acute effect on lipolysis – 3T3-F442A DIFFERENTIATION 48h 72h NUTRITION D0 GROWTH D12 ± Isoprenalin or Caffein 0 0,5 1 1,5 2 2,5 day 12 Glycerol[µM] Glycerol release in BIOMIMESYS®Adipose tissue 0 µM 1 µM 10 µM 0 0,5 1 1,5 2 2,5 day 12 Glycerol[µM] Glycerol release in BIOMIMESYS®Adipose tissue 0 mM 5 mM 10 mM Preadipocytes and adipocytes cultured in BIOMIMESYS®Adipose tissue can be used as models for lipogenesis and lipolysis analysis. 90mins Isoprenalin treatment Caffein treatment +50% +60% Adipocyte differentiation 0 10 20 30 40 50 60 day7 day14 day28 RFUAdipoRed/DNA Lipids accumulation in BIOMIMESYS® Adipose tissue without RA 1µM RA 5µM RA 10µM RA ** *** *** *** *** *** *** BIOMIMESYS®Adipose tissue BIOMIMESYS®Adipose tissue 0 1 2 3 4 5 6 7 8 day1 day7 day14 day21 day28 FABP4/RPII FABP4 mRNA expression 2D BIOMIMESYS®Adipocytes ** *** *** BIOMIMESYS®Adipose tissue 0 5 10 15 20 25 30 35 40 45 50 day1 day7 day14 day21 day28 RFUAdipoRed/DNA 3T3-L1 2D BIOMIMESYS®AdipocyteBIOMIMESYS®Adipose tissue 0 0,2 0,4 0,6 0,8 1 1,2 day7 day14 day21 day28 GLUT-4/RPII GLUT-4 mRNA expression 2D BIOMIMESYS®AdipocyteBIOMIMESYS®Adipose tissue

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