Modulation of mesenchymal stromal cells properties by the microenvironment in 3D culture

Authors

DOI:

https://doi.org/10.15587/2519-8025.2023.288082

Keywords:

mesenchymal stromal cells, three-dimensional cultivation, blood plasma hydrogel, alginate microspheres, macroporous scaffolds

Abstract

The aim of the research was to compare the shape, viability, metabolic and proliferative activity of mesenchymal stromal cells (MSCs) during cultivation in hydrogels and macroporous scaffolds.

Materials and methods. Human adipose tissue MSCs were isolated from lipoaspirates of healthy adult donors after obtaining informed consent. Hydrogels were obtained from platelet-poor human blood plasma and alginate polymer, cross-linked with calcium ions in microspheres. Macroporous scaffolds were prepared from plasma by the cryotropic gelation method. Morphology and viability of cells within carriers were assessed using vital dyes. Metabolic and proliferative activity of MSCs was studied by the Alamar Blue test on the 1st, 3rd and 7th day of 3D culturing.

Results. Three-dimensional blood plasma scaffolds had a branched pore structure with a size sufficient for cell proliferation and migration. When plasma proteins were cross-linked with L-cysteine, almost all MSCs were viable, attached to the pore surface, spread and proliferated, filling carrier cavities. In plasma hydrogels, MSCs occupied spaces and acquired a fibroblast-like morphology, maintaining viability. In alginate microspheres, MSCs were uniform distributed throughout the gel volume, kept their spherical shape, but had high viability. The highest metabolic activity of MSCs was observed in macroporous scaffolds, the lowest one in alginate microspheres. During cultivation, the activity of cells in macroporous scaffolds and plasma hydrogels increased significantly, which indirectly indicated the proliferation processes.

Conclusions. Properties of MSCs during 3D cultivation significantly depend on the microenvironment: in blood plasma carriers, cells acquire a fibroblast-like morphology and proliferate, while in alginate microspheres, they remain spherical and do not proliferate.

Supporting Agency

  • National Research Foundation of Ukraine (project No. 2021.01/0276)

Author Biographies

Oleksandr Petrenko, Institute of Problems of Cryobiology and Cryomedicine of National Academy of Sciences of Ukraine

Doctor of Biological Sciences, Professor,  Head of Department

Department of Cryobiochemistry

Head of Institute

Olena Rogulska, Institute of Problems of Cryobiology and Cryomedicine of National Academy of Sciences of Ukraine

PhD, Senior Researcher

Department of Cryobiochemistry

Natalia Trufanova, Institute of Problems of Cryobiology and Cryomedicine of National Academy of Sciences of Ukraine

PhD, Senior Researcher

Department of Cryobiochemistry

Oleh Trufanov, Institute of Problems of Cryobiology and Cryomedicine of National Academy of Sciences of Ukraine

PhD, Senior Researcher

Department of Cryobiochemistry

Oleksandra Hubenia, Institute of Problems of Cryobiology and Cryomedicine of National Academy of Sciences of Ukraine

Junior Researcher

Department of Cryobiochemistry

Olena Revenko, Institute of Problems of Cryobiology and Cryomedicine of National Academy of Sciences of Ukraine

PhD, Senior Researcher

Department of Cryobiochemistry

Daria Cherkashina, Institute of Problems of Cryobiology and Cryomedicine of National Academy of Sciences of Ukraine

PhD, Senior Researcher

Department of Cryobiochemistry

References

  1. Renesme, L., Pierro, M., Cobey, K. D., Mital, R., Nangle, K., Shorr, R., Lalu, M. M., Thébaud, B. (2022). Definition and Characteristics of Mesenchymal Stromal Cells in Preclinical and Clinical Studies: A Scoping Review. Stem Cells Translational Medicine, 11 (1), 44–54. doi: https://doi.org/10.1093/stcltm/szab009
  2. Langer, R., Vacanti, J. P. (1993). Tissue Engineering. Science, 260 (5110), 920–926. doi: https://doi.org/10.1126/science.8493529
  3. Rech, J., Getinger-Panek, A., Gałka, S., Bednarek, I. (2022). Origin and Composition of Exosomes as Crucial Factors in Designing Drug Delivery Systems. Applied Sciences, 12 (23), 12259. doi: https://doi.org/10.3390/app122312259
  4. Jensen, C., Teng, Y. (2020). Is It Time to Start Transitioning From 2D to 3D Cell Culture? Frontiers in Molecular Biosciences, 7. doi: https://doi.org/10.3389/fmolb.2020.00033
  5. Tan, L., Liu, X., Dou, H., Hou, Y. (2022). Characteristics and regulation of mesenchymal stem cell plasticity by the microenvironment – specific factors involved in the regulation of MSC plasticity. Genes & Diseases, 9 (2), 296–309. doi: https://doi.org/10.1016/j.gendis.2020.10.006
  6. Dhamecha, D., Movsas, R., Sano, U., Menon, J. U. (2019). Applications of alginate microspheres in therapeutics delivery and cell culture: Past, present and future. International Journal of Pharmaceutics, 569, 118627. doi: https://doi.org/10.1016/j.ijpharm.2019.118627
  7. Pravdyuk, A. I., Petrenko, Y. A., Fuller, B. J., Petrenko, A. Y. (2013). Cryopreservation of alginate encapsulated mesenchymal stromal cells. Cryobiology, 66 (3), 215–222. doi: https://doi.org/10.1016/j.cryobiol.2013.02.002
  8. Tarusin, D., Mazur, S., Volkova, N., Petrenko, Yu., Zaikov, V., Petrenko, A. (2016). Encapsulation of mesenchymal stromal cells in alginate microspheres. Biotechnologia Acta, 9 (4), 58–66. doi: https://doi.org/10.15407/biotech9.04.058
  9. Petrenko, Yu. A., Ivanov, R. V., Petrenko, A. Yu., Lozinsky, V. I. (2011). Coupling of gelatin to inner surfaces of pore walls in spongy alginate-based scaffolds facilitates the adhesion, growth and differentiation of human bone marrow mesenchymal stromal cells. Journal of Materials Science: Materials in Medicine, 22 (6), 1529–1540. doi: https://doi.org/10.1007/s10856-011-4323-6
  10. Elowsson, L., Kirsebom, H., Carmignac, V., Mattiasson, B., Durbeej, M. (2013). Evaluation of macroporous blood and plasma scaffolds for skeletal muscle tissue engineering. Biomaterials Science, 1 (4), 402–410. doi: https://doi.org/10.1039/c2bm00054g
  11. Rogulska, O., Petrenko, Y., Petrenko, A. (2016). DMSO-free cryopreservation of adipose-derived mesenchymal stromal cells: expansion medium affects post-thaw survival. Cytotechnology, 69 (2), 265–276. doi: https://doi.org/10.1007/s10616-016-0055-2
  12. Zuk, P. A., Zhu, M., Ashjian, P., De Ugarte, D. A., Huang, J. I., Mizuno, H. et al. (2002). Human Adipose Tissue Is a Source of Multipotent Stem Cells. Molecular Biology of the Cell, 13 (12), 4279–4295. doi: https://doi.org/10.1091/mbc.e02-02-0105
  13. Rogulska, O. Y., Trufanova, N. A., Petrenko, Y. A., Repin, N. V., Grischuk, V. P., Ashukina, N. O. et al. (2021). Generation of bone grafts using cryopreserved mesenchymal stromal cells and macroporous collagen‐nanohydroxyapatite cryogels. Journal of Biomedical Materials Research Part B: Applied Biomaterials, 110 (2), 489–499. doi: https://doi.org/10.1002/jbm.b.34927
  14. Dhurat, R., Sukesh, M. (2014). Principles and methods of preparation of platelet-rich plasma: A review and author′s perspective. Journal of Cutaneous and Aesthetic Surgery, 7 (4), 189–197. doi: https://doi.org/10.4103/0974-2077.150734
  15. Lozinsky, V. I., Galaev, I. Yu., Plieva, F. M., Savina, I. N., Jungvid, H., Mattiasson, B. (2003). Polymeric cryogels as promising materials of biotechnological interest. Trends in Biotechnology, 21 (10), 445–451. doi: https://doi.org/10.1016/j.tibtech.2003.08.002
  16. Dankberg, F., Persidsky, M. D. (1976). A test of granulocyte membrane integrity and phagocytic function. Cryobiology, 13 (4), 430–432. doi: https://doi.org/10.1016/0011-2240(76)90098-5
  17. Petrenko, Yu. A., Gorokhova, N. A., Tkachova, E. N., Petrenko, A. Yu. (2005). The reduction of Alamar Blue by peripheral blood lymphocytes and isolated mitochondria. The Ukrainian Biochemical Journal, 77 (5), 100–105.
  18. Discher, D. E., Mooney, D. J., Zandstra, P. W. (2009). Growth Factors, Matrices, and Forces Combine and Control Stem Cells. Science, 324 (5935), 1673–1677. doi: https://doi.org/10.1126/science.1171643
  19. de Vos, P., Faas, M. M., Strand, B., Calafiore, R. (2006). Alginate-based microcapsules for immunoisolation of pancreatic islets. Biomaterials, 27 (32), 5603–5617. doi: https://doi.org/10.1016/j.biomaterials.2006.07.010
  20. Kumar, V. B., Tiwari, O. S., Finkelstein-Zuta, G., Rencus-Lazar, S., Gazit, E. (2023). Design of Functional RGD Peptide-Based Biomaterials for Tissue Engineering. Pharmaceutics, 15 (2), 345. doi: https://doi.org/10.3390/pharmaceutics15020345
Modulation of mesenchymal stromal cells properties by the microenvironment in 3D culture

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Published

2023-09-30

How to Cite

Petrenko, O., Rogulska, O., Trufanova, N., Trufanov, O., Hubenia, O., Revenko, O., & Cherkashina, D. (2023). Modulation of mesenchymal stromal cells properties by the microenvironment in 3D culture. ScienceRise: Biological Science, (3(36), 28–33. https://doi.org/10.15587/2519-8025.2023.288082

Issue

No. 3(36) (2023)

Section

Biological research

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Copyright (c) 2023 Oleksandr Petrenko, Olena Rogulska, Natalia Trufanova, Oleh Trufanov, Oleksandra Hubenia, Olena Revenko, Daria Cherkashina

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