NDLI: Procedure for decellularization of porcine heart by retrograde coronary perfusion.

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Year: 2012 Issue: 70

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Procedure for decellularization of porcine heart by retrograde coronary perfusion.

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Procedure for decellularization of porcine heart by retrograde coronary perfusion.

Content Provider	World Health Organization (WHO)-Global Index Medicus
Author	Remlinger, Nathaniel T. Wearden, Peter D. Gilbert, Thomas W.
Description	Author Affiliation: Remlinger NT ( McGowan Institute for Regenerative Medicine.)
Abstract	Perfusion-based whole organ decellularization has recently gained interest in the field of tissue engineering as a means to create site-specific extracellular matrix scaffolds, while largely preserving the native architecture of the scaffold. To date, this approach has been utilized in a variety of organ systems, including the heart, lung, and liver (1-5). Previous decellularization methods for tissues without an easily accessible vascular network have relied upon prolonged exposure of tissue to solutions of detergents, acids, or enzymatic treatments as a means to remove the cellular and nuclear components from the surrounding extracellular environment(6-8). However, the effectiveness of these methods hinged upon the ability of the solutions to permeate the tissue via diffusion. In contrast, perfusion of organs through the natural vascular system effectively reduced the diffusion distance and facilitated transport of decellularization agents into the tissue and cellular components out of the tissue. Herein, we describe a method to fully decellularize an intact porcine heart through coronary retrograde perfusion. The protocol yielded a fully decellularized cardiac extracellular matrix (c-ECM) scaffold with the three-dimensional structure of the heart intact. Our method used a series of enzymes, detergents, and acids coupled with hypertonic and hypotonic rinses to aid in the lysis and removal of cells. The protocol used a Trypsin solution to detach cells from the matrix followed by Triton X-100 and sodium deoxycholate solutions to aid in removal of cellular material. The described protocol also uses perfusion speeds of greater than 2 L/min for extended periods of time. The high flow rate, coupled with solution changes allowed transport of agents to the tissue without contamination of cellular debris and ensured effective rinsing of the tissue. The described method removed all nuclear material from native porcine cardiac tissue, creating a site-specific cardiac ECM scaffold that can be used for a variety of applications.
File Format	HTM / HTML
e-ISSN	1940087X
DOI	10.3791/50059
Journal	Journal of Visualized Experiments
Issue Number	70
Language	English
Publisher	MyJove Corp.
Publisher Date	2012-12-06
Publisher Place	United States
Access Restriction	Open
Subject Keyword	Discipline Physical Sciences Discipline Life Sciences Discipline Medicine Heart Physiology Myocardial Reperfusion Myocardium Cytology Animals Swine Tissue Engineering Tissue Scaffolds Research Support, N.i.h., Extramural Video-audio Media
Content Type	Text
Resource Type	Article

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