Cryopreservation of Biological Resources
Cultural Properties of Cryopreserved Thymic Multipotent Stromal Cells and Fetal Skin and Muscle-Derived Cells
Valentina V. Nikolska Institute of Genetic and Regenerative Medicine of the National Academy of Medical Sciences of Ukraine, Kyiv
Yanina-Maria O. Semenova Institute of Genetic and Regenerative Medicine of the National Academy of Medical Sciences of Ukraine, Kyiv
Lyuba I. Taranukha Institute of Genetic and Regenerative Medicine of the National Academy of Medical Sciences of Ukraine, Kyiv
Ihor S. Nikolsky Institute of Genetic and Regenerative Medicine of the National Academy of Medical Sciences of Ukraine, Kyiv
cryopreservation, thymic multipotent stromal cells, fetal skin- and muscle-derived multipotent stromal cells, cultural properties
The paper provides a comparison of properties of cryopreserved fetal murine multipotent stromal cells (MSCs) of skin-muscular origin and those derived from adult thymus in culture in vitro. Fetal MSCs showed a 30% higher number of average population doublings within 24 hrs, and 41% lower average population doubling time. It was found that the fetal MSCs of the 4th passage had a 39% higher clonogenic activity than the adult thymus-derived ones. Fetal MSCs and those derived from adult thymus differentiated in osteogenic and adipogenic lineages with equal efficiency in special culture media. Fetal and thymus-derived MSCs were characterized by almost the same high ability of contact interaction with thymocytes, and the fibroblast-lymphocyte rosette (FLR) formation. They were far less active in FLR formation with lymph node cells. This indicated the presence of membrane affinity for immature lymphoid cells in both MSC subpopulations. The results showed the fetal MSCs to be significantly different from the adult thymus-derived MSCs by more active kinetics of growth and clonogenic potential. However, both cell subpopulations had virtually the same ability for linear differentiation and showed high activity during contact with immature lymphoid cells. Linear differentiation and the ability to interact with lymphocytes were found to be quite stable properties of MSCs, but a proliferative activity and in vitro colony formation distinguished significantly in different types of MSCs. This can be taken into account when choosing the cells for therapy, research and results assessment.
Aizenshtadt AA, Skazina MA, Kotelevskaya EA, et al. Characterization of umbilical cord mesenchymal stromal cells during long-term expansion in vitro. Herald of North-Western State Medical University named after I.I. Mechnikov. 2018;10(1):11- 9. Russian
Bifari F, Lisi V, Mimiola E, et al. Immune modulation by mesenchymal stem cells. Transfus Med Hemother. 2008; 35(3): 194-204.
Brown C, McKee C, Bakshi S, et al. Mesenchymal stem cells: cell therapy and regeneration potential. J Tissue Eng Regen Med. 2019; 13(9): 1738-55.
Freshney RJ. Culture of animal cells. Moscow: Binom. Laboratoriya Znanii; 2010. 691 p. Russian.
Goltsev AN, Dubrava TG, Kozlova YuA, et al. Investigation of the therapeutic effect of transfusion of cryopreserved hematopoietic stem cells from different sources in the treatment of experimental adjuvant arthritis. Transplantology. 2007; 9 (1): 28-30. Russian.
Goltsev AN, Rassokha IV, Lutsenko ED, et al. Intercellular interactions in immunocompetent sphere at rheumatoid arthritis following the application of hematopoietic embryonic liver cells. Problems of Cryobiology. 2003; (3): 45-53.
Gregory CA, Gunn WG, Peister A, Prockop DJ. An Alizarin red-based assay of mineralization by adherent cells in culture: comparison with cetylpyridinium chloride extraction. Anal Biochem. 2004; 329(1): 77-84.
Grishchenko VI, Lobyntseva GS, Votyakova IA, Shereshkov SI. Hematopoietic cells of the embryonic liver. Kyiv: Naukova dumka; 1988.
Grzhibovsky AM, Ivanov SV, Gorbatova MA. Analysis of quantitative data in two independent samples using Statistica and SPSS software: parametric and non-parametric tests. Science & Healthcare. 2016; (2): 5-28. Russian.
Jacobs SA, Roobrouck VD, Verfaillie CM, Van Gool SW. Immunological characteristics of human mesenchymal stem cells and multipotent adult progenitor cells. Immunol Cell Biol. 2013; 91(1): 32-9.
Kim WK, Jung H, Kim DH, et al. Regulation of adipogenic differentiation by LAR tyrosine phosphatase in human mesenchymal stem cells and 3T3-L1 preadipocytes. J Cell Sci. 2009; 122 (22): 4160-7.
Le Blanc K, Mougiakakos D. Multipotent mesenchymal stromal cells and the innate immune system. Nat Rev Immunol. 2012; 12(5): 383-96.
Nikolska KI. Peculiarities of culture and in vitro contact interaction of cryopreserved thymic multipotent stromal cells and hemopoietic cells. Probl Cryobiol Cryomed. 2018;28(1):5-13.
Nikolskaya EI, Butenko GM. Structural-functional organisation of the bone marrow hematopoietic stem cells niches. Cell and Organ Transplantology 2016; 4(1): 82-100.
Nikolsky IS, Nikolskaya VV, Zubov DA. Fibroblast and mastolymphocytic rosettes. Russian Journal of Allergy. 2011; 1 (4): 260-1. Russian.
Panchenko LM, Sych EE, Yatsenko AP. Efficiency of cloning of human bone marrow stromal stem cells in the presence of highly porous glass ceramics and its ex vivo solubility. Herald of Orthopaedics, Traumatology and Prosthetics. 2014; (4): 50-4.
Patenaude J, Perreault C. Thymic mesenchymal cells have a distinct transcriptomic profile. J Immunol. 2016; 196(11): 4760- 70.
Petrenko YA, Tarasov AI, Grischenko VI, Petrenko AY. Cryopreservation effect on immunologic activity and phenotypic composition of human embryonic liver lymphoid cells. Probl Cryobiol Cryomed. 2002; (4): 76-9.
Prockop DJ, Phinney DG, Bunnell BA. Mesenchymal stem cells: methods and protocols. Totowa: Humana Press; 2008. 192 p.
Rebrova OY. Statistical analysis of medical data: application of the Statistica software package. Moscow: Media Sfera; 2002. 312 p. Russian.
Robey P. 'Mesenchymal stem cells': fact or fiction, and implications in their therapeutic use. F1000Res. 2017; 20(6): F1000 Faculty Rev-524.
Wolfrom C, RaynaudN, Maigne J, et al. Periodic fluctuations in proliferation of SV-40 transformed human skin fibroblast lines with prolonged lifespan. Cell Biol Toxicol. 1994; 10(4): 247-54
Nikolska, V., Semenova , Y.-M., Taranukha, L., & Nikolsky, I. (2021). Cultural Properties of Cryopreserved Thymic Multipotent Stromal Cells and Fetal Skin and Muscle-Derived Cells. Problems of Cryobiology and Cryomedicine, 31(3), 249–257. https://doi.org/10.15407/cryo31.03.249