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Comparison of cardiac stem cell sheets detached by Versene solution and from thermoresponsive dishes reveals similar properties of constructs
Authors:Konstantin V. Dergilev  Pavel I. Makarevich  Zoya I. Tsokolaeva  Maria A. Boldyreva  Irina B. Beloglazova  Ekaterina S. Zubkova  Mikhail Yu. Menshikov  Yelena V. Parfyonova
Affiliation:1. Russian Cardiology Research and Production Complex, Laboratory of Angiogenesis, 121552, Moscow, Russian Federation;2. Lomonosov Moscow State University, Medical Research and Education Centre, Institute of Regenerative Medicine, Laboratory of gene and cell therapy, 119192, Moscow, Russian Federation;3. Lomonosov Moscow State University, Faculty of Medicine, Laboratory of gene and cell technologies, 119192, Moscow, Russian Federation
Abstract:Cell sheets (CS) from c-kit+ cardiac stem cell (CSC) hold a potential for application in regenerative medicine. However, manufacture of CS may require thermoresponsive dishes, which increases cost and puts one in dependence on specific materials. Alternative approaches were established recently and we conducted a short study to compare approaches for detachment of CS from c-kit+ CSC. Our in-house developed method using chelation by Versene solution was compared to UpCell? thermoresponsive plates in terms of CSC proliferation, viability, gap junction formation and engraftment in a model of myocardial infarction. Use of Versene solution instead of thermoresponsive dishes resulted in comparable CS thickness (approximately 100 mcm), cell proliferation rate and no signs of apoptosis detected in both types of constructs. However, we observed a minor reduction of gap junction count in Versene-treated CS. At day 30 after delivery to infarcted myocardium both types of CS retained at the site of transplantation and contained comparable amounts of proliferating cells indicating engraftment. Thus, we may conclude that detachment of CS from c-kit+ CSC using Versene solution followed by mechanical treatment is an alternative to thermoresponsive plates allowing use of routinely available materials to generate constructs for cardiac repair.
Keywords:BSA  bovine serum albumin  CS  cell sheet  CSC  cardiac stem cell  ECM  extracellular matrix  EDTA  ethylenediaminetetraacetic acid  FACS  fluorescence-activated cell sorting  FBS  fetal bovine serum  FOV  field of view  IMDM  Iscove's Modified Dulbecco's Medium  MI  myocardial infarction  PBS  phosphate-buffered saline  polyNIPAM  poly(N-isopropylacrylamide)  SD  standard deviation  SEM  standard error of mean  Cell sheet  Cardiac stem cell  Myocardial infarction  Thermoresponsive dish  Tissue engineering
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