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Matrix metalloproteinases as reagents for cell isolation

Paper ID Volume ID Publish Year Pages File Format Full-Text
16739 42608 2016 15 PDF Available
Title
Matrix metalloproteinases as reagents for cell isolation
Abstract

•Recombinant MMPs were compared to bacterial collagenase for mesenchymal stem cell isolation.•MMP-12 isolated cells as efficiently as bacterial collagenase with a purer population of cells.•A single activation cocktail can be used for MMPs prior to stem cell isolation.

Cell isolation methods for therapeutic purposes have seen little advancement over the years. The original methods of stem cell and islet isolation using bacterial collagenases were developed in the early 1980s and are still used today. Bacterial collagenases are subject to autodegradation, and isolates obtained with these enzymes may be contaminated with endotoxins, reducing cell viability and contributing to toxicity in downstream applications. Here we describe a novel method for isolation of mesenchymal stem cells from adipose tissue (ADSC) utilizing recombinantly produced matrix metalloproteases (MMPs). The ADSCs isolated by MMPs displayed essentially identical morphological and phenotypical characteristics to cells isolated by bacterially-derived collagenase I and Liberase™. Samples isolated with MMPs and Liberase™ had comparable levels of CD73, CD90, and CD105. The adipogenic and osteogenic potential of the ADSCs isolated by MMPs was retained as compared to cells isolated with Liberase™. However, ADSCs isolated by Liberase™ displayed 6% contamination with other cells as per negative markers revealed by PE staining, as opposed to <1% for all MMP-treated samples. MMP-based cell isolation may contribute to optimization of transplantation technology.

Keywords
Matrix metalloproteinase; E. coli; Recombinant enzyme; Clostridium histolyticum collagenase; Adipose tissue; Stem cell
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Publisher
Database: Elsevier - ScienceDirect
Journal: Enzyme and Microbial Technology - Volumes 93–94, November 2016, Pages 29–43
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
100% Money Back Guarantee
Full-text PDF Download
Online Support
Any Questions? feel free to contact us