Preparation of an adipogenic hydrogel from subcutaneous adipose tissue

Poon, Christopher; Cotta, Maria V. Pereira E.; Sinha, Shiba; Palmer, Jason; Woods, Alan A.; Morrison, Wayne A.; Abberton, Keren M.

doi:10.1016/j.actbio.2012.11.003

Cited by 71 publications

(78 citation statements)

References 54 publications

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“…Human adipose tissue has been shown to have a Young’s modulus of roughly 2 kPa [33], and thus we fabricated polyacrylamide gels using a constant 8% acrylamide monomer solution with varying bis-acrylamide monomer ranging from 0.03% to 0.5% to produce gel substrates with a stiffness range of 2 to 40 kPa (Figure 2) that remained constant across the surface of each gel. Since previous studies have shown the importance of adipose specific ECM cues, these gels were then functionalized with adipose matrix to further recapitulate the adipose microenvironment [18, 19]. After functionalization, ASCs readily adhered to the polyacrylamide gel surface (Supplementary Figure 1).…”

Section: Resultsmentioning

confidence: 99%

“…Within just the past 4 years, several groups have developed methods to extract ECM components from human adipose tissue. Presenting this complex combination of adipose-specific proteins has already been seen to foster the development of ASCs toward an adipogenic lineage [18, 19, 34]. We hoped to expand on these results by not only recreating the biochemical cues of the adipose microenvironment, but also simulate the biomechanical properties of the tissue as well.…”

Section: Discussionmentioning

confidence: 98%

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Stimulation of adipogenesis of adult adipose-derived stem cells using substrates that mimic the stiffness of adipose tissue

et al. 2013

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Biochemical and biomechanical extracellular matrix (ECM) cues have recently been shown to play a role in stimulating stem cell differentiation towards several lineages, though how they combine to induce adipogenesis has been less well studied. The objective of this study was to recapitulate both the ECM composition and mechanical properties of adipose tissue in vitro to stimulate adipogenesis of human adipose-derived stem cells (ASCs) in the absence of exogenous adipogenic growth factors and small molecules. Adipose specific ECM biochemical cues have been previously shown to influence adipogenic differentiation; however, the ability of biomechanical cues to promote adipogenesis has been less defined. Decellularized human lipoaspirate was used to functionalize polyacrylamide gels of varying stiffness to allow the cells to interact with adipose-specific ECM components. Culturing ASCs on gels that mimicked the native stiffness of adipose tissue (2 kPa) significantly upregulated adipogenic markers, in the absence of exogenous adipogenic growth factors and small molecules. As substrate stiffness increased, the cells became more spread, lost their rounded morphology, and failed to upregulate adipogenic markers. Together these data imply that as with other lineages, mechanical cues are capable of regulating adipogenesis in ASCs.

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Section: Resultsmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 98%

Stimulation of adipogenesis of adult adipose-derived stem cells using substrates that mimic the stiffness of adipose tissue

et al. 2013

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“…Proteins and glycoproteins can be extracted using a homogenization process involving pestle and mortar or high speed shear mixed within a high salt buffer that physically disrupts the ECM particles and collagen fiber structure at physiologic pH [43–47]. Homogenization involves a dispase enzymatic step that cleaves fibronectin, collagen IV, and collagen I and digests the ECM, a urea extraction step which further disrupts the non-covalent bonding and increases the solubility of the ECM proteins, and centrifugation that removes any residual non-soluble ECM components.…”

Section: Ecm Hydrogel Formationmentioning

confidence: 99%

“…Cell behavior in response to ECM hydrogels has consistently been shown to be comparable to Matrigel and/or collagen substrate for liver [87, 88], skeletal muscle [58], heart [58] and fat [43–45, 47, 67] applications. Uriel et al [43] showed that primary rat pre-adipocytes cultured on the surface of adipose ECM hydrogels (1 mg/mL) formed colonies that were significantly larger compared to Matrigel (1 mg/mL) after 7 days indicative of enhanced pre-adipocyte differentiation.…”

Section: Cellular Response To Ecm Hydrogelsmentioning

confidence: 99%

“…In general, ECM hydrogels have been well-tolerated in a wide variety of in vivo applications. No adverse immune response was shown after ECM hydrogels were injected in the heart [55, 60, 75–81], fat [43, 45, 47, 50, 67], liver [57], brain [21, 22, 24] skeletal muscle [23, 63, 64, 83], tendon [26, 59, 84], spinal cord [82], lung [49], cartilage [70], or colon [51, 71], and these studies included both homologous and heterologous ECM hydrogels. The findings in vivo are consistent with in vitro studies that have shown the pepsin-digested ECM (“pre-gel”) promotes a regulatory (“M2-like”) macrophage activation state, which is associated with a constructive remodeling response in vivo [71, 90, 91].…”

Section: In Vivo Applications Of Ecm Hydrogelsmentioning

confidence: 99%

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Extracellular matrix hydrogels from decellularized tissues: Structure and function

et al. 2017

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Extracellular matrix (ECM) bioscaffolds prepared from decellularized tissues have been used to facilitate constructive and functional tissue remodeling in a variety of clinical applications. The discovery that these ECM materials could be solubilized and subsequently manipulated to form hydrogels expanded their potential in vitro and in vivo utility; i.e. as culture substrates comparable to collagen or Matrigel, and as injectable materials that fill irregularly-shaped defects. The mechanisms by which ECM hydrogels direct cell behavior and influence remodeling outcomes are only partially understood, but likely include structural and biological signals retained from the native source tissue. The present review describes the utility, formation, and physical and biological characterization of ECM hydrogels. Two examples of clinical application are presented to demonstrate in vivo utility of ECM hydrogels in different organ systems. Finally, new research directions and clinical translation of ECM hydrogels are discussed.

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A biomimetic hydrogel functionalized with adipose ECM components as a microenvironment for the 3D culture of human and murine adipocytes

Louis¹,

Pannetier²,

Souguir³

et al. 2017

Biotech & Bioengineering

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The lack of relevant in vitro models for adipose tissue makes necessary the development of a more physiological environment providing spatial and chemical cues for the effective maturation of adipocytes. We developed a biofunctionalized hydrogel with components of adipose extracellular matrix: collagen I, collagen VI, and the cell binding domain of fibronectin and we compared it to usual 2D cultures on plastic plates. This scaffold allowed 3D culture of mature adipocytes from the preadipocytes cell lines 3T3-L1 and 3T3-F442A, as well as primary Human White Preadipocytes (HWP), acquiring in vivo-like organization, with spheroid shaped adipocytes forming multicellular aggregates. The size of these aggregates increased with time up to 120 μm in diameter after 4 weeks of maturation, with good viability. Significantly higher lipogenic activity (up to 20-fold at day 28 for HWP cultures) and differentiation rates were also observed compared to 2D. Gene expression analyses highlighted earlier differentiation and complete maturation of 3D HWP compared to 2D, reinforced by the expression of Perilipin protein after 21 days of nutrition. This increase in adipocytes phenotypic and genotypic markers made this scaffold-driven culture as a robust adipose 3D model. Retinoic acid inhibition of lipogenesis in HWP or isoprenalin and caffeine induction of lipolysis performed on mouse 3T3-F442A cells, showed higher doses of molecules than typically used in 2D, underlying the physiologic relevance of this 3D culture system. Biotechnol. Bioeng. 2017;114: 1813-1824. © 2017 Wiley Periodicals, Inc.

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Preparation of an adipogenic hydrogel from subcutaneous adipose tissue

Cited by 71 publications

References 54 publications

Stimulation of adipogenesis of adult adipose-derived stem cells using substrates that mimic the stiffness of adipose tissue

Stimulation of adipogenesis of adult adipose-derived stem cells using substrates that mimic the stiffness of adipose tissue

Extracellular matrix hydrogels from decellularized tissues: Structure and function

A biomimetic hydrogel functionalized with adipose ECM components as a microenvironment for the 3D culture of human and murine adipocytes

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