Stem cell transplantationEvaluation of Bone Marrow Mesenchymal Stem Cell Standard Cryopreservation Procedure Efficiency
Section snippets
Materials and Methods
All experiments were performed in accordance with principles of treatment of laboratory animals of the Brazilian College of Experiments in Animals (COBEA). Ten male Wistar rats (250 to 300 g) were euthanized for collection of bone marrow.
Results
Viability determinations by the trypan blue exclusion test showed 94.76% and 90.58% viability before versus after cryopreservation, while the values with the flow cytometry assay (annexin V conjugated with 7-amino-actinomycin) were 85.52% and 66.25%, respectively (Fig 1, Table 1).
Discussion
The standard cryopreservation procedure did not appear to be efficient for BMMeSCs. The flow cytometry assay was more sensitive than the trypan blue exclusion test to demonstrate viability loss of BMMeSCs. Significant cell death could be explained by shrinkage with decreased nucleus to cytoplasm ratio with adherence characteristics, which could increase the risk of freezing damage. The trypan blue exclusion test is acceptable for the determination of the integrity of the plasma membrane before
Acknowledgments
We would like to thank Faculdades Pequeno Príncipe for support.
References (12)
Adult stem cells: assessing the case for pluripotency
Trends Cell Biol
(2002)- et al.
Stem cell transplantation in myocardial infarction
Rev Cardiovasc Med
(2004) - et al.
Fundamental cryobiology of human hematopoietic progenitors cells I: osmotic characteristics and volume distribution
Cryobiology
(1998) - et al.
Measurement of the affinity and cooperativity of annexin V-membrane binding under conditions of low membrane occupancy
Anal Biochem
(2004) - et al.
Transdifferentiation of human peripheral blood CD34+-enriched cell population into cardiomyocites, endothelial cells and smooth muscle cells in vivo
Circulation
(2003) - et al.
Cellular cardiomyoplasty for myocardial regeneration
Asian Cardiovasc Thorac Ann
(2005)
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