Hypothermic Storage of Mesenchymal Stromal Cell-based Spheroids at a Temperature of 22°C
Keywords:
spheroids, mesenchymal stromal cells, hypothermic storage, actin filaments, viability, metabolic activityAbstract
Spheroids formed from mesenchymal stromal/stem cells (MSCs) provide optimal microenvironment for cell potential and stemmness support being also promising structures for relevant research models as well as cell therapies development. Practical application requires the development of effective methods for storing spheroids. Storage at ambient temperature (22°C) can be safer and more convenient for short term transportation of MSC-based spheroids. Herein we report that the spheroids formed by human adipose tissue-derived MSCs can be stored at ambient temperature (22°C) for up to 7 days with efficient preservation of viability, metabolic activity, ability to attach, and differentiation capacity. Cytoskeleton reorganization, cell size reduction, and metabolic activity decrease were shown to result from spheroidal formation and were extremely important for the development of spheroid resistance to the subsequent ambient storage.
Probl Cryobiol Cryomed 2024; 34(2):186–200
References
Alvarez-Pérez J, Ballesteros P, Cerdán S. Microscopic images of intraspheroidal pH by 1H magnetic resonance chemical shift imaging of pH sensitive indicators. MAGMA. 2005; 18(6): 293-301. CrossRef
Bahsoun S, Coopman K, Akam EC. Quantitative assessment of the impact of cryopreservation on human bone marrow-derived mesenchymal stem cells: up to 24 h post-thaw and beyond. Stem Cell Res Ther [Internet]. 2020 Dec 14 [cited 2024 Sep 10]; 11(1): 540. Available from: https://stemcellres.biomedcentral.com/articles/10.1186/s13287-020-02054-2 CrossRef
Bara JJ, Richards RG, Alini M, et al. Concise review: Bone marrow-derived mesenchymal stem cells change phenotype following in vitro culture: implications for basic research and the clinic. Stem Cells. 2014; 32(7): 1713-23. CrossRef
Campagnoli C, Roberts IA, Kumar S, et al. Identification of mesenchymal stem/progenitor cells in human first-trimester fetal blood, liver, and bone marrow. Blood. 2001; 98(8): 2396-402. CrossRef
Chatterjee A, Saha D, Niemann H, et al. Effects of cryopreservation on the epigenetic profile of cells. Cryobiology. 2017; 74: 1-7. CrossRef
Chen Y, Yu B, Xue G, et al. Effects of storage solutions on the viability of human umbilical cord mesenchymal stem cells for transplantation. Cell Transplant. 2013; 22(6): 1075-86. CrossRef
Cheng NC, Chen SY, Li JR, et al. Short-term spheroid formation enhances the regenerative capacity of adipose-derived stem cells by promoting stemness, angiogenesis, and chemotaxis. Stem Cells Transl Med. 2013; 2(8): 584-94. CrossRef
Chinnadurai R, Garcia MA, Sakurai Y, et al. Actin cytoskeletal disruption following cryopreservation alters the biodistribution of human mesenchymal stromal cells in vivo. Stem Cell Reports. 2014; 3(1): 60-72. CrossRef
Corwin WL, Baust JM, Baust JG, et al. Characterization and modulation of human mesenchymal stem cell stress pathway response following hypothermic storage. Cryobiology. 2014; 68(2): 215-26. CrossRef
Curcio E, Salerno S, Barbieri G, et al. Mass transfer and metabolic reactions in hepatocyte spheroids cultured in rotating wall gas-permeable membrane system. Biomaterials. 2007; 28(36): 5487-97. CrossRef
Dankberg, F, Persidsky, MD. A test of granulocyte membrane integrity and phagocytic function. Cryobiology. 1976; 13 (4): 430-2. CrossRef
Davies OG, Smith AJ, Cooper PR, et al. The effects of cryopreservation on cells isolated from adipose, bone marrow and dental pulp tissues. Cryobiology. 2014; 69(2): 342-7. CrossRef
Eleuteri S, Fierabracci A. Insights into the Secretome of Mesenchymal Stem Cells and Its Potential Applications. Int J Mol Sci [Internet]. 2019 Sep 17 [cited 2024 Sep 2]; 20(18): 4597. Available from: https://www.mdpi.com/1422-0067/20/18/4597 CrossRef
Erices AA, Allers CI, Conget PA, et al. Human cord bloodderived mesenchymal stem cells home and survive in the marrow of immunodeficient mice after systemic infusion. Cell Transplant. 2003; 12(6): 555-61. CrossRef
Foty R. A simple hanging drop cell culture protocol for generation of 3D spheroids. J Vis Exp [Internet]. 2011 May 6 [cited 2024 Sep 21]; (51): 2720. Available from: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3197119/ CrossRef
García-Sánchez D, Fernández D, Rodríguez-Rey JC, et al. Enhancing survival, engraftment, and osteogenic potential of mesenchymal stem cells. World J Stem Cells. 2019; 11(10): 748-63. CrossRef
Ge J, Guo L, Wang S, et al. The size of mesenchymal stem cells is a significant cause of vascular obstructions and stroke. Stem Cell Rev Rep. 2014; 10(2): 295-303. CrossRef
Gordiyenko OI, Kovalenko IF, Rogulska OY, et al. Theory-based cryopreservation mode of mesenchymal stromal cell spheroids. Cryobiology [Internet]. 2024 Jun [cited 2024 Oct 9]; 115:104906. Available from: https://www.sciencedirect.com/science/article/abs/pii/S0011224024000610 CrossRef
Guo L, Zhou Y, Wang S, et al. Epigenetic changes of mesenchymal stem cells in three-dimensional (3D) spheroids. J Cell Mol Med. 2014; 18(10): 2009-19. CrossRef
Jiang B; Yan L; Miao Z; et al. Spheroidal formation preserves human stem cells for prolonged time under ambient conditions for facile storage and transportation. Biomaterials 2017; 133: 275-86. CrossRef
Le Blanc K, Ringdén O. Immunobiology of human mesenchymal stem cells and future use in hematopoietic stem cell transplantation. Biol Blood Marrow Transplant. 2005; 11(5):321-34. CrossRef
Li Y, Dong Y, Ran Y, et al. Three-dimensional cultured mesenchymal stem cells enhance repair of ischemic stroke through inhibition of microglia. Stem Cell Res Ther [Internet]. 2021 Jun 21 [cited 2024 Sep 22]; 12(1): 358 Available from: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8218508/ CrossRef
Liang X, Ding Y, Zhang Y, et al. Paracrine mechanisms of mesenchymal stem cell-based therapy: current status and perspectives. Cell Transplant. 2014; 23(9): 1045-59. CrossRef
Liu Y, Muñoz N, Tsai AC, et al. Metabolic Reconfi guration Supports Reacquisition of Primitive Phenotype in Human Mesenchymal Stem Cell Aggregates. Stem Cells. 2017; 35(2): 398-410. CrossRef
Murphy KC, Hung BP, Browne-Bourne S, et al. Measurement of oxygen tension within mesenchymal stem cell spheroids. J R Soc Interface [Internet]. 2017 Feb [cited 2024 Sep 12]; 14(127): 20160851. Available from: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5332570/ CrossRef
Pattappa G, Heywood HK, de Bruijn JD, et al. The metabolism of human mesenchymal stem cells during proliferation and differentiation. J Cell Physiol. 2011. 226(10): 2562-70. CrossRef
Petrenko YA, Gorokhova NA, Tkachova EN, et al. The reduction of Alamar Blue by peripheral blood lymphocytes and isolated mitochondria. Ukr Biokhim Zh. 2005; 77(5): 100-5. PubMed
Pittenger MF, Mackay AM, Beck SC, et al. Multilineage potential of adult human mesenchymal stem cells. Science. 1999; 284(5411): 143-7. CrossRef
Rovere M, Reverberi D, Arnaldi P, et al. Spheroid size influences cellular senescence and angiogenic potential of mesenchymal stromal cell-derived soluble factors and extracellular vesicles. Front Bioeng Biotechnol [Internet]. 2023 Dec 12 [cited 2024 Sep 18]; 11: 1297644. Available from: https://www.frontiersin.org/journals/bioengineering-and-biotechnology/articles/10.3389/fbioe.2023.1297644/full CrossRef
Sotirov R, Kostadinova M, Pashova S, et al. Morphology of Mesenchymal Stem Cells in 3D spheroids. Acta morphologica et anthropologica. 2018; 25: 90-6.
Tsai AC, Liu Y, Yuan X, et al. Compaction, fusion, and functional activation of three-dimensional human mesenchymal stem cell aggregate. Tissue Eng Part A. 2015; 21(9-10): 1705-19. CrossRef
Wiehe JM, Kaya Z, Homann JM, et al. GMP-adapted overexpression of CXCR4 in human mesenchymal stem cells for cardiac repair. Int J Cardiol. 2013; 167(5): 2073-81. CrossRef
Wu YD, Li M, Liao X, et al. Effects of storage culture media, temperature and duration on human adipose-derived stem cell viability for clinical use. Mol Med Rep. 2019; 19(3): 2189-201. CrossRef
Yoshino M, Kajiya M, Yoshii H, et al. Distinctive biological properties between mesenchymal stem cell spheroids and clumps of mesenchymal stem cells/extracellular matrix complexes in 3D culture systems. Appl Sci [Internet]. 2023 Nov 29 [cited 2024 Sep 20]; 13(23): 12790. Available from: https://www.mdpi.com/2076-3417/13/23/12790 CrossRef
Zhang Q, Nguyen AL, Shi S, et al. Three-dimensional spheroid culture of human gingiva-derived mesenchymal stem cells enhances mitigation of chemotherapy-induced oral mucositis. Stem Cells Dev. 2012; 21(6): 937-47. CrossRef
Zhou Y, Chen H, Li H, et al. 3D culture increases pluripotent gene expression in mesenchymal stem cells through relaxation of cytoskeleton tension. J Cell Mol Med. 2017; 21(6): 1073-84. CrossRef
Zuk PA, Zhu M, Ashjian P, et al. Human adipose tissue is a source of multipotent stem cells. Mol Biol Cell. 2002; 13(12): 4279-95. CrossRef
Downloads
Published
How to Cite
Issue
Section
License
This work is licensed under a Creative Commons Attribution 4.0 International License.
Authors who publish with this journal agree to the following terms:
- Authors retain copyright and grant the journal right of first publication with the work simultaneously licensed under a Creative Commons Attribution License that allows others to share the work with an acknowledgement of the work's authorship and initial publication in this journal.
- Authors are able to enter into separate, additional contractual arrangements for the non-exclusive distribution of the journal's published version of the work (e.g., post it to an institutional repository or publish it in a book), with an acknowledgement of its initial publication in this journal.
- Authors are permitted and encouraged to post their work online (e.g., in institutional repositories or on their website) prior to and during the submission process, as it can lead to productive exchanges, as well as earlier and greater citation of published work (See The Effect of Open Access).
