Comparative analysis of highly defined proteases for the isolation of adipose tissue-derived stem cells

Regenerative Medicine
Linda PilgaardVladimir Zachar

Abstract

Before the potential of adipose tissue-derived stem cells can fully be exploited for a broad scope of tissue-engineering and cell-based therapeutical applications, an effective and reproducible method for isolation is needed. To comparatively analyze five highly defined protease formulations, Blendzyme 1-4, liberase H1 and a crude collagenase mixture in the course of digestion that consisted of three 1-h intervals. The resulting digests of human adipose tissue aspirates were evaluated for the yield of nucleated cells, viability and frequency of specific lineages, in particular CD90, CD34 and CD45, by flow cytometry. The functionality of the cells was assessed as to the colony-forming capacity in limiting dilution assays. Based on all evaluation criteria, Blendzymes 1 and 2 and liberase H1 demonstrated a superior performance and highest consistency. Blendzyme 3 clearly underperformed compared with all other enzymes, and the performance of the rest of enzymes appeared erratic. As for the length of digestion, a 2-h interval appeared optimal when weighing both the yield and functionality of the cells in the stromal vascular fractions obtained from different adipose tissue samples. Our results demonstrate that the highly purified pr...Continue Reading

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Citations

Jun 29, 2012·BMC Physiology·Kristin J Speaker, Monika Fleshner
Oct 11, 2012·PloS One·Jeppe Grøndahl RasmussenTrine Fink
Oct 6, 2014·Journal of Plastic, Reconstructive & Aesthetic Surgery : JPRAS·Naghmeh NaderiIain S Whitaker
Mar 19, 2011·Expert Opinion on Biological Therapy·Vladimir ZacharTrine Fink
Apr 24, 2010·Journal of Biomedicine & Biotechnology·Zhiqiang LiuChangyong Wang
Nov 17, 2009·Journal of Hepatology·Laurent DolléAlbert Geerts
Jul 6, 2014·Biochemical and Biophysical Research Communications·Jens Isak AndersenCristian Pablo Pennisi
Aug 16, 2016·BioImpacts : BI·Mohammad Amir AmirkhaniMohammad Ali Nilforoushzadeh
Jan 14, 2017·Journal of Tissue Engineering and Regenerative Medicine·Joris A van DongenMartin C Harmsen
Jul 19, 2013·American Journal of Physiology. Cell Physiology·Marie-Hélène PerruchotIsabelle Louveau
Jan 11, 2012·Colloids and Surfaces. B, Biointerfaces·Steffan FoldbergVladimir Zachar

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Methods Mentioned

BETA
enzymatic dissociation
flow cytometry

Software Mentioned

FlowJo
SPSS

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