Decellularization and recellularization of organs and tissues

A decellularization, organ technology, applied in the field of organs and tissues, which can solve the problem of no ideal matrix structure, no display of organs or tissues, etc.

Active Publication Date: 2012-09-26
RGT UNIV OF MINNESOTA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the matrices developed so far often do not have the ideal matrix structure and / or have not been shown to effectively recreate the vascular bed of an organ or tissue

Method used

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  • Decellularization and recellularization of organs and tissues
  • Decellularization and recellularization of organs and tissues
  • Decellularization and recellularization of organs and tissues

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0057] Example 1 - Preparation of solid organs for decellularization

[0058] To avoid postmortem thrombus formation, donor rats were systemically heparinized with a dose of 400 U of heparin / kg of donor. After heparinization, carefully dissect the heart and adjacent great vessels.

[0059] The heart was placed in physiological saline solution (0.9%) containing heparin (2000 U / ml) and stored at 5°C until further processing. Under sterile conditions, separate the connective tissue from the heart and great vessels. The inferior vena cava and left and right pulmonary veins distal to the right and left atria were ligated with non-absorbable monofilament ligatures.

Embodiment 2

[0060] Example 2 - Cannulation and perfusion of solid organs

[0061] The heart was fixed on a decellularization device for perfusion ( figure 1 ). Cannulation of the descending thoracic artery for retrograde coronary (arterial) perfusion ( figure 1 , intubation A). Ligate branches of the thoracic arteries (eg, brachiocephalic, left common carotid, left subclavian). Cannulate the pulmonary artery before it divides into left and right pulmonary arteries ( figure 1 , intubation B). Cannulation of the superior vena cava ( figure 1 , intubation C). This configuration enables both retrograde and antegrade coronary (arterial) perfusion.

[0062] Perfusion from the coronary arteries through the capillary bed to the coronary venous system to the right atrium and superior vena cava (C) occurs when positive pressure is applied to the aortic cannula (A). Perfusion from the right atrium, coronary sinus, and coronary veins through the capillary bed to the coronary and aortic cann...

Embodiment 3

[0063] Example 3 - Decellularization

[0064] After fixation of the heart in the decellularization device, antegrade perfusion was initiated with cold heparinized calcium-depleted phosphate buffer containing 1-5 mmol adenosine / L infusion to reestablish continuous coronary (arterial) flow. Coronary (arterial) flow was assessed by measuring coronary (arterial) infusion pressure and flow, and coronary resistance was calculated. After 15 minutes of steady coronal flow, detergent-based decellularization was initiated.

[0065] The specific processing method is as follows. However, in brief, the heart is perfused antegradely with detergent. After perfusion, flush the heart retrogradely with buffer (eg, PBS). Then, the hearts were perfused with PBS containing antibiotics, and then with PBS containing DNase I. The heart was then perfused with 1% benzalkonium chloride to reduce and prevent future microbial contamination, and the organ was perfused with PBS to remove any residual ...

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Abstract

The invention provides for methods and materials to decellularize a solid organ and to recellularize such a decellularized organ to thereby generate a solid organ.

Description

technical field [0001] The present invention relates to organs and tissues, and more particularly to methods and materials for decellularization and recellularization of organs and tissues. Background technique [0002] Biologically derived matrices have been studied in the fields of tissue engineering and regeneration. However, the matrices developed so far often do not have the ideal matrix structure and / or have not been shown to effectively recreate the vascular bed of an organ or tissue. The present invention describes methods of decellularizing and recellularizing organs and tissues. Summary of the invention [0003] The present invention provides methods and materials for decellularizing an organ or tissue, and methods and materials for recellularizing a decellularized organ or tissue. [0004] In one aspect, the invention provides a decellularized mammalian heart. The decellularized mammalian heart includes an extracellular matrix having an outer surface of the d...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61M1/10C12N5/06A61F2/06A61K35/34
CPCA61K35/34C12M21/08C12M25/14C12M33/04C12N5/0657C12N2533/90A61P43/00A61K9/0024C12N5/0602C12N2506/00C12M21/14A61M60/00A61K35/12
Inventor H·奥特D·泰勒
Owner RGT UNIV OF MINNESOTA
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