INFLUENCE OF THE STERILIZATION METHOD ON THE PROLIFERATION AND VIABILITY OF MOLT-4 CELLS ENCAPSULATED IN ALGINATE GEL
https://doi.org/10.17650/1726-9784-2020-19-3-46-51
Abstract
Introduction. Alginate gel encapsulation is widely used in biomedical research to protect cells from mechanical stress and contact with the recipient’s immune system. The use of alginate for transplanting biomedical cellular products to humans imposes high requirements on the sterility of the final product. To select the optimal protocol for cell encapsulation for further in vitro use or transplantation of cells enclosed in an alginate gel, it is necessary to study how sterilization processes affect such characteristics of the obtained gel as the ability to maintain cell viability and proliferation.
The purpose of the study was to compare the effect of ultraviolet radiation and autoclaving on the biological properties of alginate gel.
Materials and methods. MOLT-4 culture cells were encapsulated in 1, 2 and 4 % alginate beads at a concentration of 0.75 × 106 cells/ml. On the eighth day of cultivation, we measured the size of the colonies and determined the viability of the cells in an automatic cell counter.
Results. The colony area for all variants of the concentration of alginate gel was significantly higher for a gel sterilized by UV treatment compared with autoclaving. The difference between the colony sizes for the two methods of alginate sterilization was significant at the accepted level of significance for all variants of gel concentrations (α = 0.05, df = 198, t = 1.972). Moreover, the number of viable cells after dissolution of alginate did not significantly differ between the experimental variants.
Conclusion. Thus, autoclaving sterilization leads to proliferation retardation of encapsulated cells and cannot be recommended for use in protocols for the preparation of alginate capsules with encapsulated cells for in vitro and in vivo cultivation.
About the Authors
O. I. Kit
National Medical Research Center of Oncology of the Ministry of Health of the Russian Federation
Russian Federation
Russian Federation
S. Yu. Filippova
National Medical Research Center of Oncology of the Ministry of Health of the Russian Federation
Russian Federation
Russian Federation
A. O. Sitkovskaya
National Medical Research Center of Oncology of the Ministry of Health of the Russian Federation
Russian Federation
Russian Federation
V. V. Pozdnyakova
National Medical Research Center of Oncology of the Ministry of Health of the Russian Federation
Russian Federation
Russian Federation
O. V. Khokhlova
National Medical Research Center of Oncology of the Ministry of Health of the Russian Federation
Russian Federation
Russian Federation
References
1. Omami M., McGarrigle J.J., Reedy M. et al. Islet microencapsulation: strategies and clinical status in diabetes. Curr Diab Rep 2017;17(7):47. DOI: 10.1007/s11892-017-0877-0.
2. Espona-Noguera A., Ciriza J., CañibanoHernández A. et al. Review of advanced hydrogel-based cell encapsulation systems for insulin delivery in type 1 diabetes mellitus. Pharmaceutics 2019;11(11):597. DOI: 10.3390/pharmaceutics11110597.
3. Cernigliaro V., Peluso R., Zedda B. et al. Evolving cell-based and cell-free clinical strategies for treating severe human liver diseases. Cells 2020;9(2):386. DOI: 10.3390/cells9020386.
4. Bhere D., Shah K. Stem Cell-Based Therapies for Cancer. Adv Cancer Res 2015;127:159–89. DOI: 10.1016/bs.acr.2015.04.012.
5. Mohr A., Zwacka R. The future of mesenchymal stem cell-based therapeutic approaches for cancer – from cells to ghosts. Cancer Lett 2018;414:239–49. DOI: 10.1016/j.canlet.2017.11.025.
6. Andersen T., Auk-Emblem P., Dornish M. 3D Cell Culture in Alginate Hydrogels. Microarrays (Basel) 2015;4(2):133–61. DOI: 10.3390/microarrays4020133.
7. Read T., Sorensen D.R., Mahesparan R. et al. Local endostatin treatment of gliomas administered by microencapsulated producer cells. Nat Biotechnol 2001;19(1):29–34. DOI: 10.1038/83471.
8. Teng H., Zhang Y., Wang W. et al. Inhibition of tumor growth in mice by endostatin derived from abdominal transplanted encapsulated cells. Acta Biochim Biophys Sin (Shanghai) 2007;39(4):278–84. DOI: 10.1111/j.1745-7270.2007.00273.x.
9. Zhang Y., Wang W., Xie Y. et al. In vivo culture of encapsulated endostatinsecreting Chinese hamster ovary cells for systemic tumor inhibition. Hum Gene Ther 2007;18(5):474–81. DOI: 10.1089/hum.2006.166.
10. Hodder E., Duin S., Kilian D. et al. Investigating the effect of sterilisation methods on the physical properties and cytocompatibility of methyl cellulose used in combination with alginate for 3D-bioplotting of chondrocytes. J Mater Sci Mater Med 2019;30(1):10. DOI: 10.1007/s10856-018-6211-9.
Review
For citations:
Kit O.I., Filippova S.Yu., Sitkovskaya A.O., Pozdnyakova V.V., Khokhlova O.V. INFLUENCE OF THE STERILIZATION METHOD ON THE PROLIFERATION AND VIABILITY OF MOLT-4 CELLS ENCAPSULATED IN ALGINATE GEL. Russian Journal of Biotherapy. 2020;19(3):46-51. (In Russ.) https://doi.org/10.17650/1726-9784-2020-19-3-46-51
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