Biopolym. Cell. 2022; 38(2):93-102.
Molecular and Cell Biotechnologies
Obtaining and characterization of rat fetuses neural cells aggregates/spheroids
1, 2Sukach O. M., 1Vsevolodska S. O., 1Ochenashko O. V., 1Shevchenko M. V.
- Institute for Problems of Cryobiology and Cryomedicine, NAS of Ukraine
23, Pereyaslavskaya Str., Kharkiv, Ukraine, 61015 - G. S. Skovoroda Kharkiv National Pedagogical University
29, Alchevskyh Str., Kharkiv, Ukraine, 61002
Abstract
Aim. Search for optimal conditions for the formation of aggregates/spheroids by neural cells (NC) isolated from the brain tissue of rat fetuses. Study of behavioral characteristics of NC in culture as a part of three-dimensional aggregates/spheroids. Methods. Isolation of NC from fetal rat brain tissue; cell viability assessment; cultivation; immunocytochemical staining; NC proliferative activity assessment. Results. It was found that the conditions for the robust formation of aggregates are: 1) the presence of 10 % adult rat’s blood serum in cell culture medium, 2) the seeding cell density – 1–4×106 cells/ml and 3) the initial cell viability higher than 20 %. Conclusions. The isolated rat fetuses brain cells, when cultured in the presence of 10 % serum and at concentrations above 1×106 cells/ml, spontaneously form multicellular aggregates. The structure of aggregates formed during short-term cultivation depends on the initial cell viability, and the size of aggregates depends on the initial viability and seeding densities of NC. The rat fetuses NC aggregates/spheroids contain stem/progenitor cells that can proliferate and differentiate. NC aggregates/spheroids form three-dimensional structures, in which favorable conditions are developed for the survival and adequate functioning of the committed and stem/progenitor NC.
Keywords: rat fetuses, neural cells, aggregates, spheroids, cultivation.
Full text: (PDF, in English)
References
[1]
Neurological Disorders: Public Health Challenges, W. H. Organization, Geneva, 2006. 218 p.
[2]
Yano S, Miwa S, Mii S, Hiroshima Y, Uehara F, Kishimoto H, Tazawa H, Zhao M, Bouvet M, Fujiwara T. Cancer cells mimic in vivo spatial-temporal cell-cycle phase distribution and chemosensitivity in 3-dimensional Gelfoam histoculture but not 2-dimensional culture as visualized with real-time FUCCI imaging. Cell Cycle. 2015; 14(6):808-19.
[3]
Her GJ, Wu HC, Chen MH, Chen MY, Chang SC, Wang TW. Control of three-dimensional substrate stiffness to manipulate mesenchymal stem cell fate toward neuronal or glial lineages. Acta Biomater. 2013; 9(2):5170-80.
[4]
Bozza A, Coates EE, Incitti T, Ferlin KM, Messina A, Menna E, Bozzi Y, Fisher JP, Casarosa S. Neural differen-tiation of pluripotent cells in 3D alginate-based cultures. Biomaterials. 2014; 35(16):4636-45.
[5]
Daud MF, Pawar KC, Claeyssens F, Ryan AJ, Haycock JW. An aligned 3D neuronal-glial co-culture model for peripheral nerve studies. Biomaterials. 2012; 33:5901-13.
[6]
Sukach OM, Shevchenko MV. Three-dimensional cell cultivation systems. Biopolym Cell. 2020; 36(3):182-96.
[7]
Petrenko AYu, Sukach AN. Isolation of intact mitochondria and hepatocytes using vibration. Anal Biochem. 1991; 194(2): 326-9.
[8]
Sukach AN, Shevchenko MV, Liashenko TD. Comparative study of influence on fetal bovine serum and serum of adult rat on cultivation of newborn rat neural cells. Biopolym Cell. 2014; 30(5):394-400.
[9]
Coder DM. Assessment of Cell. In: Robinson JP, editor. Current Protocols in Cytometry. New York: John Wiley and Sons Inc; 1997; Suppl. 15:9.2.1-9.2.14.
[10]
Lin RZ, Chou LF, Chien CC, Chang HY. Dynamic analysis of hepatoma spheroid formation: roles of E-cadherin and β1-integrin. Cell Tissue Res. 2006; 324(3):411-22.
