Cryopreservation of Multicellular Spheroids Derived From Newborn Piglet Adrenal Glands
Application of special in vitro culture techniques for the cells, derived from different animal and human organs, makes
possible the obtaining of multicellular spheroids (MSs), being the natural 3-D environment for cells unlike the standard culture with the
cells in monolayer. Previously we have shown that MSs formed in the newborn piglet adrenal cell culture are capable to produce the
neuroblast-like cells, expressing the neuronal marker β-III-tubulin. In the present work we have assessed the regimens for MSs
cryopreservation using 5, 7 and 10% dimethyl sulfoxide (DMSO) and 25% fetal bovine serum (FBS). Analysis of some characteristics
of cryopreserved MSs (surface adhesion, capability to produce the neuroblast-like cells and monolayer formation by fibroblast-like
cells) allowed to choose the cryopreservation regimen with 1°C/min cooling rate in the presence of 10% DMSO as the most optimal
one. The FBS supplement to the medium did not significantly affect the cryopreservation outcome, although there was found a
tendency to increase the capability of cryopreserved MSs to produce the neuroblast- and fibroblast-like cells.
Probl Cryobiol Cryomed 2017; 27(4): 322-333
Bondarenko T.P., Legach E.I., Kiroshka V.V. et al. Cultivation, cryopreservation and tissue transplantation of endocrine glands. Current problems of cryobiology and cryomedicine / Ed. By A.N. Goltsev. – Kharkiv; 2012. p. 361–401.
Bozhok G.A., Sidorenko O.S., Plaksina E.M. et al. Neural differentiation potential of sympathoadrenal progenitors derived from fresh and cryopreserved neonatal porcine adrenal glands. Cryobiology 2016; 73(2): 152–161. CrossRef PubMed
Chen J., Hersmus N., Van Duppen V. et al. The adult pituitary contains a cell population displaying stem/progenitor cell and early embryonic characteristics. Endocrinology 2005; 146(9): 3985–3998. CrossRef PubMed
Ehrhart F., Schulz J.C., Katsen-Globa A. et al. A comparative study of freezing single cells and spheroids: towards a new model system for optimizing freezing protocols for cryobanking of human tumours. Cryobiology 2009; 58(2): 119–127. CrossRef PubMed
Freshney R. Culture of Animal Cells: a Manual of Basic Technique. New York: Alan R. Liss, Inc.; 1987. PubMed
Gil-Perotin S., Duran-Moreno M., Cebrian-Silla A. et al. Adult neural stem cells from the subventricular zone: a review of the neurosphere assay. Anat Rec (Hoboken) 2013; 296(9): 1435–1452. CrossRef PubMed
Hammarback J.A., Palm S.L., Furcht L.T., Letourneau P.C. Guidance of neurite outgrowth by pathways of substratum-adsorbed laminin. Journal of Neuroscience Research 1985; 13(1–2): 213–220. CrossRef PubMed
Koenigsmann M.P., Koenigsmann M., Notter M. et al. Adhesion molecules on peripheral blood-derived CD34+ cells: effects of cryopreservation and short-term ex vivo incubation with serum and cytokines. Bone Marrow Transplant 1998; 22(11): 1077–1085. CrossRef PubMed
Kuzmuk K., Schook L. Pigs as a model for biomedical sciences. In: Rothschild M.F., Ruvinsky A., editors. The Genetics of the Pig, 2nd ed. Cambridge: Oxfordshire, 2011. p. 426–444. CrossRef
Martin-Ibanez R., Hovatta O., Canals J. Cryopreservation of human pluripotent stem cells: are we going in the right direction? In: Katkov I.I., editor. Current Frontiers in Cryobiology. Croatia: InTech, 2012. p. 139–165. CrossRef
Morris C.B. Cryopreservation of animal and human cell lines. In: Day J.G., McLellan M.R., editors. Methods in Molecular Biology, Cryopreservation and Freeze-drying Protocols. New Jersey: Humana Press Inc., 1995; p. 179–187. CrossRef
Nyberg S.L., 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(8): 901–910. CrossRef PubMed
Plaksina E.M., Sidorenko O.S., Legach E.I. et al. Expression of β- III-tubulin in the neonatal adrenal cell culture: comparison of monolayer and 3D-culture. Vestnik Khar'kovskogo natsional'nogo universiteta imeni V. N. Karazina, Seriya 'Biologiya' 2017; 28: 76-86.
Sanie-Jahromi F., Ahmadieh H., Soheili Z.S. et al. Enhanced generation of retinal progenitor cells from human retinal pigment epithelial cells induced by amniotic fluid. BMC Res Notes 2012; 5 (182). CrossRef PubMed
Santana M.M., Chung K.F., Vukicevic V. et al. Isolation, characterization, and differentiation of progenitor cells from human adult adrenal medulla. Stem Cells Transl Med 2012; 1(11): 783–791. CrossRef PubMed
Saxena S., Wahl J., Huber-Lang M.S. et al. Generation of murine sympathoadrenergic progenitor-like cells from embryonic stem cells and postnatal adrenal glands. PLoS One 2013; 8(5): e64454. CrossRef PubMed
Sidorenko O.S., Bozhok G.A., Legach E.I., Bondarenko T.P. Formation of cytospheres and neuronal differentiation in newborn piglet adrenal cell culture. Probl Cryobiol Cryomed 2013; 23(4): 359–362.
Su G.H. Pancreatic cancer: Methods and protocols. Totowa, New Jersey: Humana Press Inc.; 2005.
Sukach A.N. Influence of DMSO on the viability of human embryonic nerve cells and their behavior under in vitro culture conditions. Probl Cryobiol 2005; 15(3): 429–432.
Tamarina I., Bozhok G., Gurina T. et al. Cryopreservation of newborn mice adrenal cell suspension II. Effect caused by concentration of serum as component of cryoprotective medium. Probl Cryobiol Cryomed 2013; 23(1): 66–74.
Tan F.C., Lee K.H., Gouk S.S. et al. Optimization of cryopreservation of stem cells cultured as neurospheres: comparison between vitrification, slow–cooling and rapid cooling freezing protocols. CryoLetters 2007; 28(6): 445–460.
Xiong H., Gendelman H.E. Current Laboratory Methods in Neuroscience Research. New York: Springer-Verlag; 2014. CrossRef
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