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Proteomics Clin Appl. 2016 Jan;10(1):75-83. doi: 10.1002/prca.201500048. Epub 2015 Sep 28.

Quantification of decellularized human myocardial matrix: A comparison of six patients.

Author information

1
Department of Bioengineering, Sanford Consortium for Regenerative Medicine, University of California at San Diego, La Jolla, CA, USA.
2
Department of Biochemistry and Molecular Genetics, School of Medicine, University of Colorado, Aurora, CO, USA.
3
Department of Pediatrics (Cardiology), University of California San Diego and Rady Children's Hospital, San Diego, La Jolla, CA, USA.
4
Division of Cardiovascular Diseases, Mayo Clinic, Rochester, MN, USA.
#
Contributed equally

Abstract

PURPOSE:

The purpose of this study was to characterize and quantitatively analyze human cardiac extracellular matrix (ECM) isolated from six different cadaveric donor hearts.

EXPERIMENTAL DESIGN:

ECM was isolated by decellularization of six human cadaveric donor hearts and characterized by quantifying sulfated glycosaminoglycan content (sGAG) and via PAGE. The protein content was then quantified using ECM-targeted Quantitative conCATamers (QconCAT) by LC-SRM analysis using 83 stable isotope labeled (SIL) peptides representing 48 different proteins. Nontargeted global analysis was also implemented using LC-MS/MS.

RESULTS:

The sGAG content, PAGE, and QconCAT proteomics analysis showed significant variation between each of the six patient samples. The quantitative proteomics indicated that the majority of the protein content was composed of various fibrillar collagen components. Also, quantification of difficult to remove cellular proteins represented less than 1% of total protein content, which is very low for a decellularized biomaterial. Global proteomics identified over 200 distinct proteins present in the human cardiac ECM.

CONCLUSION AND CLINICAL RELEVANCE:

In conclusion, quantification and characterization of human myocardial ECM showed significant patient-to-patient variability between the six investigated patients. This is an important outcome for the development of allogeneic derived biomaterials and for increasing our understanding of human myocardial ECM composition.

KEYWORDS:

Decellularization; Extracellular matrix; MS; QconCAT

PMID:
26172914
PMCID:
PMC4703489
DOI:
10.1002/prca.201500048
[Indexed for MEDLINE]
Free PMC Article

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