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Bioengineered tissue constructs and cardiac uses thereof

A technology for constructs and hearts, applied in the field of implantation or attachment of cell-matrix constructs, implantation or attachment of bioengineered tissue constructs, and promotion of angiogenesis in tissues and organs

Inactive Publication Date: 2009-05-13
ORGANOGENESIS
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Preparation of tissue equivalents without supporting components or scaffolds leads to scientific challenges in creating new bioengineered tissues

Method used

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  • Bioengineered tissue constructs and cardiac uses thereof
  • Bioengineered tissue constructs and cardiac uses thereof
  • Bioengineered tissue constructs and cardiac uses thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0153] Example 1: Formation of Collagen Matrix by Human Neonatal Foreskin Fibroblasts

[0154] Human neonatal foreskin fibroblasts (derived from Organogenesis, Inc.Canton, MA) were 5×10 5 cell / 162cm 2Inoculate into tissue culture treated flasks (Costar Corp., Cambridge, MA, cat # 3150) and grow in growth medium. The growth medium consisted of Dulbecco's modified Eagle's medium (DMEM) supplemented with 10% newborn calf serum (NBCS) (HyClone Laboratories, Inc., Logan, Utah) and 4 mM L-glutamine (BioWhittaker, Walkersville, MD). ) (high glucose formulation, without L-glutamine, BioWhittaker, Walkersville, MD). Keep these cells at 37±1°C, 10±1% CO 2 atmosphere in the incubator. The medium was replaced with freshly prepared medium every 2-3 days. After 8 days in culture, cells have grown to confluence, that is, cells have formed a packed monolayer along the bottom of the tissue culture flask, and the culture medium is aspirated from the flask. Sterile filtered phosphate buffe...

Embodiment 2

[0165] Example 2: Full thickness skin constructs

[0166] Using the dermal construct formed using the method described in Example 1, normal human neonatal foreskin epidermal keratinocytes (derived from Organogenesis, Inc. Canton, MA) were seeded on the cell-matrix construct to form the surface of the skin construct. cortex.

[0167] Aseptically remove the medium from the culture insert and its surroundings. Normal human epidermal keratinocytes were expanded from frozen subcultured cell stocks to passage 4 to confluence. Cells were then detached from the dish using trypsin-verene, mixed, centrifuged to form a cell pellet, resuspended in epidermalization medium, counted and divided into 4.5 × 10 4 cells / cm 2 The density is seeded on top of the membrane. Then at 37±1℃, 10% CO 2 The constructs were incubated for 90 minutes at 90°C to allow keratinocytes to attach. After incubation, the constructs were submerged in epidermalization medium. The epidermization medium consists ...

Embodiment 3

[0171] Example 3: In vitro formation of a collagen matrix by human neonatal foreskin fibroblasts in a chemically defined medium

[0172] Using the method described in Example 1, human neonatal foreskin fibroblasts were expanded. Then resuspend the cells to 3 x 10 6 The concentration of cells / ml is 3.0×10 6 cell / TW(6.6×10 5 cells / cm 2 ) on 0.4 micron pore size, 24 mm diameter tissue culture treated membrane inserts in 6-well trays. These cells were then maintained as in Example 1 with medium omitting neonatal calf serum throughout. More specifically, the medium contained: DMEM, a 3:1 matrix mixture of HamsF-12 medium (Quality Biologics, Gaithersburg, MD), 4 mM GlutaMAX (Gibco BRL, Grand Island, NY) and additives: 5 ng / mL human recombinant Epidermal growth factor (Upstate Biotechnology, Lake Placid, NY), 0.4 μg / ml hydrocortisone (Sigma St.Louis, MO), 1×10 -4 M ethanolamine (Fluka, Ronkonkoma, NY cat. #02400 ACS grade), 1 x 10 -4 M o-phosphoryl-ethanolamine (Sigma, St. Lou...

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Abstract

Cultured tissue constructs comprising cultured cells and endogenously produced extracellular matrix components without the requirement of exogenous matrix components or network support or scaffold members. Some tissue constructs of the invention are comprised of multiple cell layers or more than one cell type. The tissue constructs of the invention have morphological features and functions similar to tissues their cells are derived and their strength makes them easily handleable. Preferred cultured tissue constructs of the invention are prepared in defined media, that is, without the addition of chemically undefined components. These tissue constructs are used to repair cardiac tissues.

Description

[0001] The invention belongs to the field of tissue engineering. The present invention relates to the implantation or attachment of bioengineered tissue constructs to promote endothelialization and vascularization in the heart and related tissues. Background of the invention [0002] Coronary heart disease is a leading cause of death in the United States today (American Heart Association's "1999 Heart and Stroke Update Statistics"). The disease, like various other cardiovascular disorders, is characterized by narrowing of the arteries and insufficient blood flow to critical tissues. [0003] Currently used clinical methods for improving blood flow in a diseased or otherwise damaged heart include invasive surgical techniques such as coronary artery bypass surgery, angioplasty and endarterectomy. Such procedures naturally involve a high degree of inherent risk during and after surgery, often providing only temporary treatment of myocardial ischemia. [0004] In an effort to im...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61F2/04
CPCA61L27/3633A61L27/3895A61B2017/00575A61B2017/00597A61B17/12122A61B2017/00592A61B2017/00606A61B17/00491A61B2017/00893A61L27/3804A61L27/3839A61B17/12172A61B17/0057A61B2017/1205A61B2017/00831A61L31/125A61L31/16A61L2430/36
Inventor P·R·比尔波D·C·埃克隆德
Owner ORGANOGENESIS
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