The analysis of the proliferative activity of cells on microparticles obtained from decellularized liver and kidney tissue
https://doi.org/10.15825/1995-1191-2018-4-69-75
Abstract
Aim. To develop the protocols for liver and kidney tissue decellularization, and to develop an analysis of the proliferative activity of human Hep-G2 hepatocarcinoma cells on various carriers.
Materials and methods. Decellularization of the liver and kidneys was performed by perfusion of detergent solutions with gradually increasing concentrations of Triton X-100 (1, 2 and 3%). A histological analysis of the obtained samples was performed, and the method of optical and scanning electron microscopy was used to study the obtained samples. The proliferative activity of human Hep-G2 hepatocarcinoma cells was studied on the obtained samples of decellularized liver and kidney tissue.
Results. Decellularization of the organ does not lead to changes in the specific structure of the tissue matrix. Microparticles with an average size of 200 μm were made from their decellularized matrix of liver and kidney tissues. The level of proliferative activity of human Hep-G2 hepatocarcinoma cells cultured on microparticles from a decellularized liver was significantly higher than on microparticles from a decellularized kidney.
Conclusion. The decellularized matrix retains the native three-dimensional structure of the tissue. The level of cell proliferative activity is significantly higher on microparticles from the decellularized liver, which confirms the preservation of the specificity of the extracellular matrix of the tissue after the process of decellularization.
Supporting agencies: the Russian Scientific Foundation (Project No. 177510098)
About the Authors
M. M. Bobrova
V.I. Shumakov National Medical Research Center of Transplantology and Artificial Organs of the Ministry of Healthcare of the Russian Federation; Lomonosov Moscow State University
Russian Federation
Russian Federation
Laboratory of Bionanotechnology, V.I. Shumakov National Medical Research Center of Transplantology and Artificial Organs of the Ministry of Healthcare of the Russian Federation; Biological faculty, Lomonosov Moscow State University
Moscow
L. A. Safonova
V.I. Shumakov National Medical Research Center of Transplantology and Artificial Organs of the Ministry of Healthcare of the Russian Federation; Lomonosov Moscow State University
Russian Federation
Russian Federation
Laboratory of Bionanotechnology, V.I. Shumakov National Medical Research Center of Transplantology and Artificial Organs of the Ministry of Healthcare of the Russian Federation; Biological faculty, Lomonosov Moscow State University
Moscow
O. I. Agapova
V.I. Shumakov National Medical Research Center of Transplantology and Artificial Organs of the Ministry of Healthcare of the Russian Federation
Russian Federation
Russian Federation
Laboratory of Bionanotechnology
Moscow
A. E. Efimov
V.I. Shumakov National Medical Research Center of Transplantology and Artificial Organs of the Ministry of Healthcare of the Russian Federation
Russian Federation
Russian Federation
Laboratory of Bionanotechnology
Moscow
I. I. Agapov
V.I. Shumakov National Medical Research Center of Transplantology and Artificial Organs of the Ministry of Healthcare of the Russian Federation
Russian Federation
Russian Federation
Laboratory of Bionanotechnology
1, Shchukinskaya str., Moscow, 123182.
Теl. (985) 231-60-42.
References
1. Nari GA, Cid M, Comín R et al. Preparation of a threedimensional extracellular matrix by decellularization of rabbit livers. Rev Esp Enferm Dig. 2013; 105 (3): 138–143.
2. Ye JS, Stoltz JF, Isla N, Liu Y, Yin YF, Zhang L. An approach to preparing decellularized whole liver organ scaffold in rat. Biomed Mater Eng. 2015; 25: 159–166.
3. Nakayama KH, Batchelder CA, Lee GE, Tarantal AF. Decellularized rhesus monkey kidney as a three-dimensional scaffold for renal tissue engineering. Tissue Eng Part A. 2010; 16 (7): 2207–2216.
4. Lutolf MP, Hubbell JA. Synthetic biomaterials as instructive extracellular microenvironments for morphogenesis in tissue engineering. Nat Biotechnol. 2005; 23 (1): 47–55.
5. Lee JS; Cho SW. Liver tissue engineering: Recent advances in the development of a bio-artificial liver. Biotechnol. Bioprocess. Eng. 2012; 17 (3): 427−438.
6. SotoGutierrez A, Zhang L, Medberry C et al. A Whole-Organ regenerative medicine approach for liver replacement. Tissue Eng Part C Methods. 2011; 17 (6): 677–686.
7. Nelson CM, Bissell MJ. Of extracellular matrix, scaffolds, and signaling: tissue architecture regulates development, homeostasis, and cancer. Annu Rev Cell Dev Biol. 2006; 22: 287–309.
8. Barnes CA, Brison J, Michel R et al. The surface molecular functionality of decellularized extracellular matrices. Biomaterials. 2011; 32 (1): 137–143.
9. Hussein KH, Park KM, Teotia PK. Fabrication of a biodegradable xenoantigen-free rat liver scaffold for potential drug screening applications. Transplantation Proc. 2013; 45 (8): 3092–3096.
10. Badylak SF, Freytes DO, Gilbert TW. Extracellular matrix as a biological scaffold material: structure and functional. Acta Biomater. 2009; 5 (1): 1–13.
11. Bobrova MM, Safonova LA, Agapova OI, Krasheninnikov ME, Shagidulin My, Agapov II. Liver tissue decellularization as a promising porous scaffold processing technology for tissue engineering and regenerative medicine. Sovrem. Technol Med. 2015; 7 (4): 6–13.
12. Mossman T. Rapid colorometric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. J Immunol Methods. 1983; 65 (1–2): 55–63.
13. Baptista PM, Siddiqui MM, Lozier G, Rodriquez SR, Atala A, Soker S. The use of whole organ decellularization for the generation of a vascularized liver organoid. Hepatology. 2011; 53 (2): 604–617.
14. Shupe T, Williams M, Brown A, Willenberg B, Petersen BE. Method of the decellularization of intact rat liver. Organogenesis. 2010; 6 (2): 134–136.
15. Macchiarini P, Jungebluth P, Go T et al. Clinical transplantation of a tissue-engineered airway. Lancet. 2008; 372 (9655): 2023–2029.
16. Zhou P, Lessa N, Estrada D et al. Decellularized liver matrix as a carrier for the transplantation of human fetal and primary hepatocytes in Mice. Liver Transpl. 2011; 17 (14): 418–427.
17. Crapo PM, Gilbert TW, Badylak SF. An overview of tissue and whole organ decellularization processes. Biomaterials. 2011; 32 (12): 3233–3243.
18. ArenasHerrera JE, Ko IK, Atala A, Yoo JJ. Decellularization for whole organ bioengineering. Biomed Mater. 2013; 8 (1): 014106.
19. Wang Y, Bao J, Wu Q et al. Method for perfusion decellularization of porcine whole liver and kidney for use as a scaffold for clinical-scale bioengineering engrafts. Xenotransplantation. 2015; 22 (1): 48–61.
20. Gilbert TW, Sellaro TL, Badylak SF. Decellularization of tissues and organs. Biomaterials. 2006; 27 (19): 3675–3683.
21. Ren H, Shi X, Tao L et al. Evaluation of two decellularization methods in the development of an organ decellularized rat liver scaffold. Liver Int. 2013; 33 (3): 448–458.
22. Pei M, Li JT, Shoukry M, Zhang Y. A review of decellularized stem cell matrix: a novel cell expansion system for cartilage tissue engineering. European Cell Mater. 2011; 22: 333–343.
23. Uygun BE, SotoGutierrez A, Yagi H et al. Organ reengineering through development of a transplantable recellularized liver graft using decellularized liver matrix. Nat Med. 2010; 16 (7): 814–820.
24. Canning P, Tan L, Chu K, Lee SW, Gray NS, Bullock AN. Structural mechanisms determining inhibition of the collagen receptor DDR1 by selective and multi-targeted type II kinase inhibitors. J Mol Biol. 2014; 426 (13): 2457–2470.
25. Londono R, Badylak SF. Biological scaffolds for regenerative medicine: mechanisms of in vivo remodeling. Ann Biomed Eng. 2015; 43 (3): 577–592.
26. Shirakigawa N, Ijima H, Takei T. Decellularized liver as a practical scaffold with a vascular network template for liver tissue engineering. J Biosci Bioeng. 2012; 114 (5): 546–551.
Review
For citations:
Bobrova M.M., Safonova L.A., Agapova O.I., Efimov A.E., Agapov I.I. The analysis of the proliferative activity of cells on microparticles obtained from decellularized liver and kidney tissue. Russian Journal of Transplantology and Artificial Organs. 2018;20(4):69-75. https://doi.org/10.15825/1995-1191-2018-4-69-75
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