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Decellularized adipose tissue

A technique for adipose tissue and decellularization, used in tissue regeneration, tissue culture, animal cells, etc.

Inactive Publication Date: 2013-02-13
QUEENS UNIV OF KINGSTON
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In general, pure natural scaffold materials are more popular, but there is a potential disadvantage in use, that is, pure natural materials need to undergo processing such as decellularization before use, so that the possibility of immunogenic reactions occurs Minimize and ensure long-term stability

Method used

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  • Decellularized adipose tissue
  • Decellularized adipose tissue
  • Decellularized adipose tissue

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0098] Material

[0099] All chemicals were purchased from Sigma-Aldrich Canada (Oakville, Canada) unless otherwise stated and used as received.

[0100] adipose tissue acquisition

[0101] Ex vivo adipose tissue samples and lipoaspirates were collected from female patients undergoing elective abdominal or thoracic surgery at Kingston General Hospital and Hotel Dieu Hospital, Canada. These tissues would normally have been discarded as part of routine operating procedures. These samples were sent to the laboratory on ice and extracted for no more than 2 hours in sterile strips of cation-free phosphate-buffered saline (PBS).

[0102] Adipose Tissue Decellularization

[0103] A decellularization protocol detailed in Table 3 was used for the processing of ex vivo adipose tissue and adipose extract material. Prior to processing, freshly isolated adipose tissue samples are cut into small pieces weighing approximately 20-25 grams. As for liposuction, its volume was chosen to pro...

Embodiment 2

[0147] The decellularized adipose tissue described here was used as a culture substrate for adherent cells, and more particularly for attachment and proliferation of human adipose stem cells on decellularized adipose tissue scaffolds. The result is as Figure 8 (a) and (b), shown with CellTracker TM Green (Invitrogen)-labeled adipose stem cells (shown in white) and decellularized adipose tissue labeled with Alexa fluor 350 (Invitrogen) (shown in gray) were easily visualized under a confocal microscope. Figure 8 (a) and (b) show representative images of 1×10 6 Adipose stem cells were seeded on decellularized adipose tissue (200mg) and 2.5×10 6Adipose-derived stem cells were seeded on decellularized adipose tissue (200 mg), and the two were in hypoxia (5% O 2 , 37°C) for 7 days. Figure 8 (c) is shown using PicoGreen (Invitrogen) ADSC proliferation on decellularized adipose tissue scaffolds was analyzed by measuring total DNA content. Decellularized adipose tissue scaffo...

Embodiment 3

[0149] Preliminary in vivo studies on decellularized adipose tissue have been carried out in a subcutaneous transplantation model. The study complied with the guidelines of the Canadian Council for the Care of Animals (CCAC). All work was managed at the Animal Disposal Laboratory within Botterell Hall, Queen's University. Efforts to develop an animal care protocol were reviewed and approved by the Queen's University Animal Care Committee (UACC).

[0150] Figure 9 (a) to 9(f) show histological and microscopic data. In these studies, 1 × 10 6 A decellularized adipose tissue scaffold isolated from adipose-derived stem cells from Wistar male mice was transplanted into the dorsal dermis of Wistar female mice ( Figure 9 (a)). Each mouse received two unseeded and two decellularized adipose tissue scaffolds seeded with ADSCs. At 4, 7, and 14 days post-transplantation, the animals were humanely sacrificed, and then the decellularized adipose tissue scaffolds were removed. Macr...

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PUM

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Abstract

This invention provides a method for decellularizing adipose tissue, comprising subjecting the adipose tissue to one or more incubations in an enzymatic digestion solution containing one or more enzymes, and one or more solvent extractions, wherein decellularized adipose tissue comprising an extracellular matrix with well-preserved three-dimensional structure is obtained. The invention also provides a decellularized adipose tissue comprising an extracellular matrix with well-preserved three-dimensional architecture, and bioscaffolds. microcarrier beads, and coatings comprising the decellularized adipose tissue.

Description

[0001] related application [0002] This application claims the benefit of priority to US Provisional Patent No. 61 / 287,236, filed December 17, 2009, the entire contents of which are hereby incorporated by reference in their entirety. technical field [0003] The present invention relates to a method for decellularizing adipose tissue and decellularized adipose tissue having an extracellular matrix with a well-preserved three-dimensional structure. This decellularized adipose tissue is suitable for use in the biomedical field, for example in soft tissue augmentation or as a bioscaffold. Furthermore, this decellularized adipose tissue can also be used as a cell culture medium substrate in static culture of primary cells or cell lines in a bioreactor or in vivo. Background technique [0004] Currently, biomaterials used clinically for soft tissue filling mainly include autologous, allogeneic and heterogeneous. Research interest in autologous materials has mostly focused on t...

Claims

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Application Information

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IPC IPC(8): C12N5/077A61K35/12A61K47/46A61K8/98A61L15/40A61L27/36A61P7/04C09D189/04C12N5/071
CPCA61L2430/40C12N5/0653C12N2533/90A61L27/3683C12N11/02A61L27/3604A61P7/04A61P17/02A61P41/00A61L2300/64A61L2400/04A61L2400/06A61L27/3834A61L27/50
Inventor L·E·弗林
Owner QUEENS UNIV OF KINGSTON
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