Cell and Organ Transplantology. 2016; 4(2):200-205.
Morphological and functional characteristics of cryopreserved multipotent mesenchymal stromal cells from bone marrow, adipose tissue and tendons
Volkovа N. A., Yukhta M. S., Goltsev A. N.
Institute for Рroblems of Сryobiology and Сryomedicine NAS of Ukraine, Kharkiv, Ukraine
The aim of study was to comparatively evaluate the morphological and functional properties of cryopreserved multipotent mesenchymal stromal cells (MMSCs) from bone marrow, fat and tendon.
Materials and methods. MMSC cultures obtained from rat bone marrow, fat and tendon. The cells was cryopreserved under protection of 10 % DMSO and 20% FBS with cooling rate of 1 deg/min down to -80°C followed by plunging into liquid nitrogen. In the studied cultures the membrane integrity, immunophenotype, ability to colony formation, proliferative characteristics (MTT-test), directed differentiation and type I collagen synthesis were evaluated.
Results. Investigated cryopreserved cell culture derived from bone marrow, tendon and adipose tissue had high membrane integrity indicators, colony formation and proliferation as well as the ability to directional adipogenic and chondrogenic differentiation. The analysis of immunophenotype showed that the tested cryopreserved MMSCs culture characterized by high levels of expression (≥90 %) of CD44, CD90, CD105, CD73 and low expression (≤1 %) of hematopoietic marker CD45. Cryopreserved bone marrow MMSCs were characterized by a high content of cells that synthesized type I collagen as compared to cultures which were derived from fat and tendon.
Conclusions. Cell cultures derived from all studied sources have immunophenotype of precursor cells of mesenchymal origin. The MMSC of tendon tissue characterized by a greater capacity for colony formation and proliferation, and lower capacity for directed adipogenic differentiation, than MSCs from bone marrow and adipose tissue.
Key words: multipotent mesenchymal stromal cells; cryopreservation; CFU; cell differentiationFull Text PDF (eng) Full Text PDF (ua)
|1. Dominici M, Le Blanc K, Mueller I, et al. Minimal criteria for defining multipotent mesenchymal stromal cells. The International Society for Cellular Therapy position statement Cytotherapу. 2006; 8(4):315-17.|
|2. Baksh D, Yao R, Tuan R. Comparison of proliferative and multilineage differentiation potential of human mesenchymal stem cells derived from umbilical cord and bone marrow. Stem cells. 2007; 25(4):1384-92.
|3. Busser H, Najar M, Raicevic G, et al. Isolation and Characterization of Human Mesenchymal Stromal Cell Subpopulations: Comparison of Bone Marrow and Adipose Tissue. Stem cells and development. 2015; 24(18):2142-57.
|4. Díaz-Prado S, Mui-os-López E, Hermida-Gómez T, et al. Multilineage differentiation potential of cells isolated from the human amniotic membrane. J Cell Biochem. 2010; 111(4):846-57.
|5. Hegyi B, Sagi1 B, Kovacs J. Identical, similar or different? Learning about immunomodulatory function of mesenchymal stem cells isolated from various mouse tissues: bone marrow, spleen, thymus and aorta wall. Int Immunology. 2010; 22(7):551-59.
|6. Suzdaltsev G, Burunova VV, Vakhrushev IB. Sravnenie sposobnosti k differentsirovke v tkani mezodermal’nogo proiskhozhdeniya mezenkhimal’nykh kletok cheloveka, vydelennykh iz raznykh istochnikov [Comparison of ability to differentiate into tissue of mesodermal origin human mesenchymal cells isolated from different sources]. Kletochnye tekhnologii v biologii i meditsine – Cell Technologies in Biology and Medicine. 2007; 1:3-10. [in Russian]|
|7. Stanco D, Viganт M, Perucca C, et al. In vitro characterization of stem/progenitor cells from semitendinosus and gracilis tendons as a possible new tool for cell-based therapy for tendon disorders. Joints. 2014; 2(4):159-68.|
|8. Angelo PC, Ferreira AC, Fonseca VD, et al. Cryopreservation does not alter karyotype, multipotency, or NANOG/SOX2 gene expression of amniotic fluid mesenchymal stem cells. Genet Mol Res. 2012; 11(2):1002-12.
|9. Rui YF, Lui PP, Li G, et al. Isolation and characterization of multipotent rat tendon-derived stem cells. Tissue Eng Part A. 2010; 16:1549-58.
|10. Pittenger MF, Mackay AM, Beck SC, et al. Multilineage potential of adult human mesenchymal stem cells. Science. 1999; 284(2):143-47.
|11. Pogozhykh D, Prokopyuk V, Pogozhykh O, et al. Influence of factors of cryopreservation and hypothermic storage on survival and functional parameters of multipotent stromal cells of placental origin. PLoS One. 2015; 2(10):0139834.
|12. Volk SW, Wang Y, Hankenson KD. Effects of donor characteristics and ex vivo expansion on canine mesenchymal stem cell properties: implications for msc-based therapies. Cell Transplant. 2012; 21(10):2189-200. doi:10.3727/096368912X636821
|13. Hagmann S, Moradi B, Frank S, et al. Different culture media affect growth characteristics, surface marker distribution and chondrogenic differentiation of human bone marrow-derived mesenchymal stromal cells. BMC Musculoskelet Disord. 2013;14:223.
|14. Giordano A, Galderisi U, Marino IR. From the laboratory bench to the patient’s bedside: An update on clinical trials with mesenchymal stem cells. Journal of Cellular Physiology. 2007; 211(1):27-35.
|15. Barry FP, Murphy JM. Mesenchymal stem cells: clinical applications and biological characterization. The International Journal of Biochemistry & Cell Biology. 2004; 36(4):568-84.
|16. Garcia-Olmo D, Garcia-Arranz M, Herreros D, et al. A phase I clinical trial of the treatment of crohn’s fistula by adipose mesenchymal stem cell transplantation. Dis Colon Rectum. 2005; 48(7):1416-23.
|17. Rogulska OYu, Revenko O, Petrenko YuA, Petrenko AYu. Proliferativno-differentsirovochnyy potentsial mul’tipotentnykh mezenkhimal’nykh stromal’nykh kletok zhirovoy tkani pri kul’tivirovanii v prisutstvii trombotsitarnogo lizata [Proliferative and differentiation potential of adipose tissue-derived mesenchymal stromal cells in the presence of platelet lysate]. Geny i kletki – Genes and Cells. 2014; 9(2):63-67. [in Russian]|
|18. Gonda K, Shigeura T, Sato T, et al. Preserved proliferative capacity and multipotency of human adipose-derived stem cells after long-term cryopreservation. Plast Reconstr Surg. 2008; 121(2):401-10.
|19. Adams RLP. Cell culture for biochemists. Amsterdam and New York: Elsevier, 1980. 292 pp.|
|20. Volkova NA, Goltsev AN. Сryopreservation effect on proliferation and differentiation potential of cultured chorion cells. CryoLetters. 2015; 36(1):25-29.|
|21. Mossman T. Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. J Immunol Methods. 1983; 65:55-63.
|22. Pratheesh MD, Gade NE, Katiyar AN, et al. Isolation, culture and characterization of caprine mesenchymal stem cells derived from amniotic fluid. Isolation Res Vet Sci. 2013; 94:313-319.
|23. Volkova NO. Doslіdzhennya morfofunktsіonal’nikh kharakteristik krіokonservovanikh kul’tur klіtin stromal’nogo pokhodzhennya [Study of morphological characteristics of cryopreserved cell cultures of stromal origin]. Problemy kriobiologii – Problems of Cryobiology and Cryomedicine. 2012; 22(2):118-25. [in Ukrainian]|
|24. Sethe S, Scutt A, Stolzing A, et al. Aging of mesenchymal stem cells. Ageing Res Rev. 2006; 5(1):91-116.
|25. Haack-Sørensen M, Ekblond A, Kastrup J. Cryopreservation and Revival of Human Mesenchymal Stromal Cells. Methods Mol Biol. 2016; 1416:357-74.
Volkovа NA, Yukhta MS, Goltsev AN. Morphological and functional characteristics of cryopreserved multipotent mesenchymal stromal cells from bone marrow, adipose tissue and tendons. Cell and Organ Transplantology. 2016; 4(2):200-205. doi:10.22494/cot.v4i2.64