Optimal delipidation solvent to secure extracellular matrix from human perirenal adipose tissue

Human adipose tissue includes useful substrates for regenerative medicine such as the extracellular matrix (ECM), but most perirenal fat tissue is wasted after kidney surgery. Since a lot of adipose tissue can be procured after a kidney, we extracted ECM from human perirenal adipose tissue and optimized the extraction process. To verify the efficacy for ECM extraction, we compared the products in several steps. Perirenal adipose tissue was either finely homogenized or underwent crude manual dissection. The amount of extracted ECM was quantified with ELISA for verification of the initial tissue downsizing effect. To validate the drying effect for fast and complete delipidation, tissues were prepared in a dry or wet phase, and residual lipids were visualized with Oil-Red-O staining. The extracted lipid was assayed at each time point to quantify the appropriate delipidation time. To select the optimal decellularization method, tissues were treated with physical, chemical, or enzymatic method, and the residual cell debris were identified with histological staining. The biochemical properties of the ECM extracted by the above methods were analyzed. The ECM extracted by fine homogenization showed a significantly enhanced amount of collagen, laminin and fibronectin compared to the crude dissection method. The dried tissue showed fast and complete lipid elimination compared to the wet tissue. Complete delipidation was achieved at 45 min after acetone treatment. Additionally, 1% triton X-100 chemical treatment showed complete decellularization with well-preserved collagen fibers. Biochemical analysis revealed preserved ECM proteins, a high cell proliferation rate and normal cell morphology without cell debris or lipids. The established process of homogenization, drying, delipidation with acetone, and decellularization with Triton X-100 treatment can be an optimal method for ECM extraction from human perirenal adipose tissue. Using this technique, human perirenal adipose tissue may be a valuable source for tissue engineering and regenerative medicine.

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701-P: Effects of Exercise on Extracellular Matrix Modifiers in Subcutaneous Adipose Tissue of Obese Adults

Diabetes ◽

10.2337/db20-701-p ◽

2020 ◽

Vol 69 (Supplement 1) ◽

pp. 701-P

Author(s):

PALLAVI VARSHNEY ◽

BENJAMIN J. RYAN ◽

CHIWOON AHN ◽

MICHAEL W. SCHLEH ◽

JEFFREY F. HOROWITZ

Keyword(s):

Extracellular Matrix ◽

Adipose Tissue ◽

Subcutaneous Adipose Tissue ◽

Obese Adults ◽

Subcutaneous Adipose

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Effects of dietary roughage on organic pig performance, behaviour and antioxidants accretion in perirenal adipose tissue

Livestock Science ◽

10.1016/j.livsci.2020.104255 ◽

2020 ◽

Vol 241 ◽

pp. 104255

Author(s):

I. Argemí-Armengol ◽

D. Villalba ◽

Marc Tor ◽

Juan Ramón Bertolín ◽

M.A. Latorre ◽

...

Keyword(s):

Adipose Tissue ◽

Perirenal Adipose Tissue ◽

Pig Performance

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14 Characterization of adipose tissue extracellular matrix component mRNA expression during porcine adipogenesis using real-time PCR

Journal of Animal Science ◽

10.1093/jas/skz397.080 ◽

2020 ◽

Vol 98 (Supplement_2) ◽

pp. 35-35

Author(s):

Maegan A Reeves ◽

Courtney E Charlton ◽

Terry D Brandebourg

Keyword(s):

Extracellular Matrix ◽

Adipose Tissue ◽

Mrna Expression ◽

Real Time ◽

Real Time Pcr ◽

Collagen Type ◽

Primary Cultures ◽

Collagen Type I ◽

Type I ◽

Matrix Metallopeptidase

Abstract Given adipose tissue is histologically classified as connective tissue, we hypothesized expression of extracellular matrix (ECM) components are significantly altered during adipogenesis. However, little is known about the regulation of the ECM during adipose tissue development in the pig. Therefore, the objective of this study was to characterize expression of ECM components during porcine adipogenesis. Primary cultures of adipose tissue stromal-vascular cells were harvested from 3-day-old neonatal pigs (n=6) and preadipocytes induced to differentiate in vitro for 8 days in the presence of insulin, hydrocortisone, and rosiglitazone. Total RNA was extracted from these cultures on days 0 and 8 post-induction. Real-time PCR was then utilized to determine changes in mRNA expression for collagen type I alpha 1 chain (COL1A), collagen type I alpha 2 chain (COL2A), collagen type I alpha 3 chain (COL3A), collagen type I alpha 4 chain (COL4A), collagen type I alpha 6 chain (COL6A), biglycan, fibronectin, laminin, nitogen-1 (NID1), matrix metallopeptidase 2 (MMP2), matrix metallopeptidase 9 (MMP9), metallopeptidase inhibitor 3 (TIMP3). The mRNA abundances of COL1A, COL3A and MMP2 were significantly downregulated 2.86-fold (P < 0.05), 16.7-fold (P < 0.01) and 3.1-fold (P < 0.05) respectively in day 8 (differentiated) compared to day 0 (undifferentiated) cultures. Meanwhile, mRNA abundances were significantly upregulated during adipogenesis for the COL2A (2.82-fold; P < 0.05), COL4A (2.01-fold; P < 0.05), COL6A (2.8-fold; P < 0.05), biglycan (49.9- fold; P < 0.001), fibronectin (452-fold; P < 0.001), laminin (6.1-fold; P < 0.05), NID1(47.4-fold; P < 0.01), MMP9 (76.8- fold; P < 0.01), and TIMP3(3.04-fold; P < 0.05) genes. These data support the hypothesis that significant changes in ECM components occur during porcine adipogenesis. Modulating adipose tissue ECM remodeling might be a novel strategy to manipulate adiposity in the pig.

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Adipose tissue exposed to high fat diet affects extracellular matrix genes in the mesenchymal stem cell population

Atherosclerosis ◽

10.1016/j.atherosclerosis.2021.06.430 ◽

2021 ◽

Vol 331 ◽

pp. e144

Author(s):

U.T. Arasu ◽

T. Örd ◽

J. Liikkanen ◽

S. Kettunen ◽

T. Lonnberg ◽

...

Keyword(s):

Extracellular Matrix ◽

Adipose Tissue ◽

Stem Cell ◽

Mesenchymal Stem Cell ◽

Cell Population ◽

High Fat Diet ◽

Stem Cell Population ◽

High Fat ◽

Mesenchymal Stem Cell Population

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Effects of photoperiod and feeding level on perirenal adipose tissue metabolic activity and leptin synthesis in the ovariectomized ewe

Reproduction Nutrition Development ◽

10.1051/rnd:19980501 ◽

1998 ◽

Vol 38 (5) ◽

pp. 489-498 ◽

Cited By ~ 71

Author(s):

François Bocquier ◽

Muriel Bonnet ◽

Yannick Faulconnier ◽

Michèle Guerre-Millo ◽

Patrice Martin ◽

...

Keyword(s):

Adipose Tissue ◽

Metabolic Activity ◽

Feeding Level ◽

Perirenal Adipose Tissue

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Augmentation of Dermal Wound Healing by Adipose Tissue-Derived Stromal Cells (ASC)

Bioengineering ◽

10.3390/bioengineering5040091 ◽

2018 ◽

Vol 5 (4) ◽

pp. 91 ◽

Cited By ~ 5

Author(s):

Joris van Dongen ◽

Martin Harmsen ◽

Berend van der Lei ◽

Hieronymus Stevens

Keyword(s):

Extracellular Matrix ◽

Wound Healing ◽

Adipose Tissue ◽

Stromal Cells ◽

Randomized Clinical Trials ◽

Cell Types ◽

Matrix Remodeling ◽

Skin Wound Healing ◽

Dermal Wound Healing ◽

Dermal Wound

The skin is the largest organ of the human body and is the first line of defense against physical and biological damage. Thus, the skin is equipped to self-repair and regenerates after trauma. Skin regeneration after damage comprises a tightly spatial-temporally regulated process of wound healing that involves virtually all cell types in the skin. Wound healing features five partially overlapping stages: homeostasis, inflammation, proliferation, re-epithelization, and finally resolution or fibrosis. Dysreguled wound healing may resolve in dermal scarring. Adipose tissue is long known for its suppressive influence on dermal scarring. Cultured adipose tissue-derived stromal cells (ASCs) secrete a plethora of regenerative growth factors and immune mediators that influence processes during wound healing e.g., angiogenesis, modulation of inflammation and extracellular matrix remodeling. In clinical practice, ASCs are usually administered as part of fractionated adipose tissue i.e., as part of enzymatically isolated SVF (cellular SVF), mechanically isolated SVF (tissue SVF), or as lipograft. Enzymatic isolation of SVF obtained adipose tissue results in suspension of adipocyte-free cells (cSVF) that lack intact intercellular adhesions or connections to extracellular matrix (ECM). Mechanical isolation of SVF from adipose tissue destructs the parenchyma (adipocytes), which results in a tissue SVF (tSVF) with intact connections between cells, as well as matrix. To date, due to a lack of well-designed prospective randomized clinical trials, neither cSVF, tSVF, whole adipose tissue, or cultured ASCs can be indicated as the preferred preparation procedure prior to therapeutic administration. In this review, we present and discuss current literature regarding the different administration options to apply ASCs (i.e., cultured ASCs, cSVF, tSVF, and lipografting) to augment dermal wound healing, as well as the available indications for clinical efficacy.

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