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Optimization of cryopreservation procedures for porcine endothelial progenitor cells

Paper ID Volume ID Publish Year Pages File Format Full-Text
20799 43191 2012 7 PDF Available
Title
Optimization of cryopreservation procedures for porcine endothelial progenitor cells
Abstract

Endothelial progenitor cells (EPCs) provide a powerful option for therapeutic use in ischemic diseases. The cell therapy-induced vasculogenesis requires sufficient homogeneous cells, and cryopreservation is a prerequisite for long-term storage and quality assurance of EPCs. The aim of this study was to optimize cryopreservation protocols of EPCs derived from porcine bone marrow. Bone marrow-derived mononuclear cells (MNCs) were isolated by density centrifugation and differentiated into EPCs. The first passage EPCs were frozen by using different methodologies, and after cryopreservation the thawed cells were cultured to the fourth passage. The recovery efficiency and functions of these cells were evaluated by determination of cell viability, proliferation and migration. We found the optimal conditions for cryopreservation of EPCs as follows: (i) a cryopreservation medium consisting of 10% dimethylsulphoxide (DMSO) in combination with 50% fetal bovine serum (FBS); (ii) using a controlled freezing rate at 5°C/min; (iii) at an optimal density of 5 × 106/ml for cryopreserved EPCs; (iv) a storage temperature of − 156°C. Under these conditions we demonstrated that EPCs could be stored in mechanical freezer for up to 18 months after cryopreservation without losing their phenotypic characteristics and biological functions.

► We optimize cryopreservation protocols of EPCs derived from porcine bone marrow. ► We compare different cryopreservation methodologies by using cell viability. ► EPCs are stored 18 months without losing characteristics and decreasing functions.

Keywords
BM, bone marrow; DAB, diaminobenzidine; DMSO, dimethylsulphoxide; EPCs, endothelial progenitor cells; FBS, fetal bovine serum; IMDM, Iscove's modified Dulbecco's medium; M199, medium 199; MNCs, mononuclear cells; PBS, phosphate buffer solution; VEGF, vasc
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Optimization of cryopreservation procedures for porcine endothelial progenitor cells
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Publisher
Database: Elsevier - ScienceDirect
Journal: Journal of Bioscience and Bioengineering - Volume 113, Issue 1, January 2012, Pages 117–123
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