Dynamics of Dimethyl Sulfoxide Penetration Into L929 Cells and L929-Based Spheroids
Keywords:L929 cells, spheroids, filtration coefficients, permeability coefficients, dimethylsulfoxide, osmotically inactive volume, cryopreservation
The study proposes an algorithm for calculating of appreciable permeability coefficients for multicellular structures in a cryoprotectant medium using physical and mathematical model of mass transfer. The values of surface-area-to-volume ratio for L929 cells at different temperatures were determined and the thermal expansion coefficient of the surface area of cell membranes was calculated (β = 2.7 × 10-3 /°C). The osmotically inactive volume for L929 cells and their spheroids was determined. Filtration and permeability coefficients to DMSO for L929 cells and in toto spheroids were found from the dynamic curves of relative volume change. The calculated parameters are the highest for individual cells and significantly (p <0.05) decrease for cells in the spheroids with increasing depth of their location, this reduction may be stipulated by a decrease in the available surface of cells in the spheroids for the penetration of extracellular substances. Obtained in this research permeability characteristics of spheroids can be used to develop optimal cryopreservation regimens for them.
Probl Cryobiol Cryomed 2021; 31(4): 316–325
Abu-Absi SF, Friend JR, Hansen LK, HuW-S. Structural polarity and functional bile canaliculi in rat hepatocyte spheroids. Exp Cell Res. 2002; 274: 56-67. CrossRef
Achilli T-M, Meyer J, Morgan JR. Advances in the formation, use and understanding of multi-cellular spheroids. Expert Opin Biol Ther. 2012; 12(10): 1347-60. CrossRef
Akiyama M, Nonomura H, Kamil SH, Ignotz RA. Periosteal cell pellet culture system:a new technique for bone engineering. Cell Transplant. 2006; 15: 521-32. CrossRef
Arai K, Murata D, Takao S, et al. Cryopreservation method for spheroids and fabrication of scaffold-free tubular constructs. PLoS ONE [Internet]. 2020 Apr 02 [cited 2021 May 15]; 15(4):e0230428. Available from: https://journals.plos.org/plosone/article?id=10.1371/journal.pone.0230428 CrossRef
Frese KK; Tuveson DA. Maximizing mouse cancer models. Nat Rev Cancer. 2007; 7: 645-58. CrossRef
Gordiyenko ОІ. [Estimation of the thermal expansion coefficient of erythrocyte membrane surface by shift of erythrocyte distribution curve by spherical index]. Biophysical Bulletin. 2003; 13 (2): 78-81. Ukranian.
Gordiyenko YeО, Gordiyenko ОІ, Maruschenko VV, Sakun OV. [Improved model for the passive mass transfer through the cell plasma membrane]. Biophysical Bulletin. 2008; 21(2): 75-80. Ukranian.
Gordiyenko YeO, Pushkar NS. [Physical basis for low temperature preservation of cell suspensions]. Кyiv: Naukova dumka; 1994. 140 p. Russian.
Griffith LG, Swartz MA. Capturing complex 3D tissue physiology in vitro. Nat Rev Mol Cell Biol. 2006; 7: 211-24. CrossRef
Holtfreter J. A study of the mechanisms of gastrulation. J Exp Zool. 1944; 95: 171-212. CrossRef
Kelm JM, Timmins NE, Brown CJ, et al. Method for generation of homogeneous multicellular tumor spheroids applicable to a wide variety of cell types. Biotechnol Bioeng. 2003; 83: 173-80. CrossRef
Kosheleva NV, Efremov YM, Shavkuta BS. Cell spheroid fusion: beyond liquid drops model. Sci Rep [Internet]. 2020 Jul 28 [cited 2021 May 15]; 10:12614. Available from: https://www.nature.com/articles/s41598-020-69540-8 CrossRef
Kunz-Schughart LA, Schroeder JA, Wondrak M, et al. Potential of fibroblasts to regulate the formation of three-dimensional vessel-like structures from endothelial cells in vitro. Am J Physiol Cell Physiol. 2006; 2905: 1385-98. CrossRef
Langhans SA. Three-dimensional in vitro cell culture models in drug discovery and drug repositioning. Front Pharmacol [Internet]. 2018 Jan 23 [cited 2021 May 15]; 9:6. Available from: https://www.frontiersin.org/articles/10.3389/fphar.2018.00006/full CrossRef
Lee JH, Jung DH, Lee DH, et al. Effect of spheroid aggregation on susceptibility of primary pig hepatocytes to cryopreservation. Transplant Proc. 2012; 44: 1015-7. CrossRef
Lin R-Z, Chang H-Y. Recent advances in three-dimensional multicellular spheroid culture for biomedical research. Biotechnology J. 2008; 3: 1172-84. CrossRef
Matta SG, Wobken JD, Williams FG, Bauer GE. Pancreatic islet cell reaggregation systems: Efficiency of cell reassociation and endocrine cell topography of rat islet-like aggregates. Pancreas 1994; 9: 439-49. CrossRef
Moscona A, Moscona H. The dissociation and aggregation of cells from organ rudiments of the early chick embryo. J Anat. 1952; 86: 287-301. PubMed
Nyberg SL, Hardin J, Amiot B, et al. Rapid large-scale formation of porcine hepatocyte spheroids in a novel spheroid reservoir bioartificial liver. Liver Transpl. 2005; 11: 901-10. CrossRef
Ogurtsova VV, Kovalenko SYe, Kovalenko IF, Gordiyenko OI. Determination of osmotically inactive volume of murine enterocytes. Probl Cryobiol Cryomed. 2016; 26(1): 93-7. CrossRef
Pampaloni F, Reynaud EG, Stelzer EH. The third dimension bridges the gap between cell culture and live tissue. Nat Rev Mol Cell Biol. 2007; 8: 839-45. CrossRef
Pinto B, Henriques AC, Silva PM, Bousbaa H. Three-dimensional spheroids as in vitro preclinical models for cancer research. Pharmaceutics [Internet]. 2020 Dec 6 [cited 2021 May 15]; 12(12):1186. Available from: https://microbialcellfactories.biomedcentral.com/articles/10.1186/s12934-015-383-5 CrossRef
Ryu N-E, Lee S-H, Park H. Spheroid culture system methods and applications for mesenchymal stem cells. Cells [Internet]. 2019 Dec 12 [cited 2021 May 15]; 8:1620. Available from: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6953111/ CrossRef
Suenaga H, Furukawa KS, Suzuki Y, et al. Bone regeneration in calvarial defects in a rat model by implantation of human bone marrow-derived mesenchymal stromal cell spheroids. J Mater Sci Mater Med [Internet]. 2015 Oct 08 [cited 2021 May 15]; 26:254. Available from: https://link.springer.com/article/10.1007%2Fs10856-015-5591-3 CrossRef
Yamaguchi Y, Ohno J, Sato A, et al. Mesenchymal stem cell spheroids exhibit enhanced in-vitro and in-vivo osteoregenerative potential. BMC Biotechnol [Internet]. 2014 Dec 06 [cited 2021 May 15]; 14:105. Available from: https://bmcbiotechnol.biomedcentral.com/articles/10.1186/s12896-014-0105-9 CrossRef
How to Cite
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).