Effect of Encapsulation into Alginate Microspheres on Viability of Mesenchymal Stromal Cells after Exposure with Penetrating Cryoprotectants
Keywords:mesenchymal stromal cells, alginate microspheres, vitrification, multicomponent solution of cryoprotectants, dimethyl sulfoxide, ethylene glycol, 1, 2-propane diol
The effect of encapsulation into alginate microspheres (AMS) on viability of mesenchymal stromal cells (MSCs) after exposure to the solutions of penetrating cryoprotectants dimethyl sulfoxide (DMSO), ethylene glycol (EG) and 1,2-propane diol (1,2‑PD) with concentrations varying from 1.5 to 9 M during 1.5 and 5 min was studied. It was shown that the viability of MSCs both as a suspension and encapsulated in AMS decreased with a rise in cryoprotectant concentration and exposure time. Encapsulation into AMS significantly protected the cells from cytotoxic damage of the cryoprotectants, e. g. if the 5-min exposure to 9 M DMSO, EG and 1,2-PD resulted in the death of quite all the cells in the suspension, the viability of encapsulated MSCs after exposure under similar conditions made 60, 80 and 52% respectively. It was found that the studied penetrating cryoprotectants had a various cytotoxicity to MSCs, and it decreased in the row DMSO > 1,2-PD > EG. The findings can be used to develop a multi-component vitrifying solution.
Probl Cryobiol Cryomed 2016; 26(3): 213–220
Aye M., Di Giorgio C., De Mo M. et al. Assessment of the geno-toxicity of three cryoprotectants used for human oocyte vitrification: dimethyl sulfoxide, ethylene glycol and propylene glycol. Food and Chemical Toxicology 2010; 48(7): 1905–1912. CrossRef PubMed
Bernas T., Dobrucki J. Mitochondrial and nonmitochondrial reduction of MTT: interaction of MTT with TMRE, JC-1, and NAO mitochondrial fluorescent probes. Cytometry 2002; 47(4): 236–242. CrossRef PubMed
Fahy G. M. The relevance of cryoprotectant "toxicity" to cryobiology. Cryobiology 1986; 23(1): 1–13. CrossRef
Gautier A., Carpentier B., Dufresne M. et al. Impact of alginate type and bead diameter on mass transfers and the metabolic activities of encapsulated c3a cells in bioartificial liver applications. European Cells and Materials 2011; 21(25): 94–106. PubMed
Goedert M., Mobed-Miremadi M. Cross-linked alginate film pore size determination using atomic force microscopy and validation using diffusivity. Journal of Surface Engineered Materials and Advanced Technology 2013; 3(4): 1–12. CrossRef
Gorokhova N.A., Petrenko Yu.A., Petrenko A.Yu. Search of cryoprotective medium for vitrification of fetal fibroblastlike suspension cells. Problems of Cryobiology 2007; 17(1): 50–59.
Herskovits T.T., Gadegbeku B., Jaillet H. On the structural stability and solvent denaturation of proteins. I. Denaturation by the alcohols and glycols. J Biol Chem 1970; 245(10): 2588–2598. PubMed
Lawson A., Mukherjee I., Sambanis A. Mathematical modeling of cryoprotectant addition and removal for the cryopreservation of engineered or natural tissues. Cryobiology 2013; 64(1): 1–11. CrossRef PubMed
Marquez-Curtis A., Janowska-Wieczorek A., McGann L., J. Elliot Mesenchymal stromal cells derived from various tissues: biological, clinical and cryopreservation aspects. Cryobiology 2015; 71(2): 181–197. CrossRef PubMed
Morris C., de Wreede L., Scholten M. et al. Should the standard dimethyl sulfoxide concentration be reduced? Results of a European Group for Blood and Marrow Transplantation prospective noninterventional study on usage and side effects of dimethyl sulfoxide. Transfusion 2014; 54(10): 2514–2522. CrossRef PubMed
Petrenko A.Yu., Mazur S.P., Petrenko Yu.A. et al. Isolation and multilineage differentiation of stromal cells from human fetal and adult tissue. Transplantologiya 2007; 9(1): 218–220.
Pravdyuk A.I., Petrenko Yu.A., Volkova N.A. et al. Properties of mesenchymal stromal human cells encapsulated in alginate microbeads. Biotechnologiya 2010; 3(2): 62–70.
Rowley S., Anderson G. Effect of DMSO exposure without cryopreservation on hematopoietic progenitor cells. Bone Marrow Transplantation 1993; 11(5): 389–393. PubMed
Taniguchi M., Arikawa R., Kaedei Y. et al. Effects of cryopro-tectant agents and equilibration methods on developmental competence of porcine oocytes. Cryoletters 2011; 32(5): 410–422.
Zaikov V.S., Petrenko Yu.A., Trufanova N.A. et al. Effect of cryopreservation using slow freezing or vitrification on viability and metabolic activity of mesenchymal stromal cells encapsulated within alginate spheres with diameter of 1 mm and more. Probl Cryobiol Cryomed 2014; 24(3): 222–230.
Zaikov V.S., Petrenko Yu.Au., Trufanova N.A. et al. Vitrification of mesenchymal stromal cells in alginate microbeads. Probl Cryobiol Cryomed 2012; 22(4): 14–20.
Wusteman M., Pegg D., Robinson M. et al. Vitrification media: toxicity, permeability, and dielectric properties. Cryobiology 2002; 44(1): 24–37. CrossRef
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