Functional Activity of Rat Adrenocorticocytes after Incubation in Dimethyl Sulfoxide Based Cryoprotective Media Supplemented with Fetal Bovine Serum
Keywords:adrenal cortex cells, dimethyl sulfoxide, fetal bovine serum, aldosterone, 3beta-hydroxysteroid dehydrogenase, JC-1
Investigation of functional state of cells at various stages of cryopreservation is important for the development and improvement of freezing protocols of endocrine cells. This study assessed the effect of incubation in media with 5, 7, 10, and 15% dimethyl sulfoxide (DMSO), as well as the media with DMSO and 25% fetal bovine serum (FBS) on some indices of adrenal cortex cell suspension: viability (MTT-test), content of cells with active 3b-hydroxysteroid dehydrogenase, functionally active mitochondria (JCâ€‘1 test), as well as the level of basal and stimulated aldosterone secretion. It was shown that presence of FBS in a cryoprotective medium reduced the toxic effect of DMSO on adrenocorticocytes during incubation, furthermore in some cases incubation in media with DMSO and FBS led to the increase in the number of cells with functionally active mitochondria and cells with active steroidogenesis if compared with the suspensions with cells not exposed to cryoprotectant solution.
Probl Cryobiol Cryomed 2014; 24(1):47â€“56.
Benton L., Shan L.X., Hardy M.P. Differentiation of adult Leydig cells. J Steroid Biochem Mol Biol 1995; 53(1): 61â€“68. CrossRef
Cossarizza A., Baccarani-Contri M., Kalashnikova G. et al. A new method for the cytofluorimetric analysis of mitochondrial membrane potential using the J-aggregate forming lipophilic cation 5,5',6,6'-tetrachloro-1,1',3,3' tetraethylbenzimidazolyl-carbocyanine iodide (JC-1). Biochem Biophys Res Commun 1993; 197(1): 40â€“45. CrossRef PubMed
Ferrari M., Fornasiero M.F., Chiara M., Isetta A.M. MTT colorimetric assay for testing macrophage cytotoxic activity in vitro. J Immunol Meth 1990; 131: 165â€“172. CrossRef
Grischenko V.I., Bondarenko T.P., Legach Ye.I. et al. Procurement, cryopreservation and clinical application of newborn piglet adrenal gland organ culture in therapy of renal failure [Methodical recommendations]. Kharkiv; 2000. PubMed
Grischenko V.I., Chuyko V.A., Pushkar M.S. Cryopreservation of endocrine organ tissues and cells. Kyiv: Naukova Dumka; 1993.
Hafez M.M., Heidger P.M. Stereological study of Leydig cell density in the guinea pig testis. J Arch Androl 1986; 17(1): 1â€“7. CrossRef
Huveneers-Oorsprong M.B., Hoogenboom L.A., Kuiper H.A. The use of the MTT test for determining the cytotoxicity of veterinary drugs in pig hepatocytes. Toxicol In Vitro 1997; 11(4): 385â€“392. CrossRef
Kardak A., Leibo S.P., Devireddy R. Membrane transport properties of equine and macaque ovarian tissues frozen in mixtures of dimethylsulfoxide and ethylene glycol. J Biomech Eng 2007; 29(5): 688â€“694. CrossRef PubMed
Karpenko L.G., Gubina N.F., Schirova V.A., Kaprelyants A.S The effects of freezing and cryoprotectant exposure on adenylate cyclase activity in cell membranes of bovine thyroid gland and adrenal cortex. CryoLetters 2001; 22(4): 229â€“234.
Legach E.I., Bozhok G.A., Bondarenko T.P. Cryopreserved organ culture of endocrine glands for hormonal insufficiency treatment. Endocr Abstracts 2010; 22: 190.
Menorval M.-A., Lluis M.M., Fernandez M.L., Reigada R. Effects of dimethyl sulfoxide in cholesterol-containing lipid membranes: a comparative study of experiments in silico and with cells. PLoS One 2012; 7: e41733. CrossRef PubMed
Murthy K.S. cAMP inhibits IP3-dependent Ca2+ release by preferential activation of cGMP-primed PKG. Am. J. Physiol. Gastrointest. Liver Physiol 2001; 275(5): 1238â€“1245.
Novikova L.A., Faletrov Ya.V., Kovaleva I.E. et al. From structure and functions of steroidogenic enzymes to new technologies of gene engineering. Biochemistry 2009; 74(13): 1482â€“1504. CrossRef
Nuydens R., Novalbos J., Dispersyn G. et al. A rapid method for the evaluation of compounds with mitochondria-protective properties. J Neurosci Methods 1999; 92(1â€“2): 153â€“159. CrossRef
Pogorelyy V.K., Turov V.V., Turov A.V. Investigation of interactions between serum albumin and dimethyl sulfoxide by PMR spectra of frozen aqueous solutions. Teor Eksperim Khimiya 1989; (1): 104â€“108.
Pushkar N.S., Chuyko V.A., Utevsky A.M. et al. Preservation and transplantation of endocrine organs and tissues. Kharkov; 1981.
Sidorenko O.S., Bozhok G.A., Legach E.I., Gurina T.M. Study of possibility to obtain and cryopreserve adrenal cell primary culture of newborn piglets. Problems of Cryobiology 2011; 21(1): 58â€“67.
Silani V. Cryopreservation of human fetal adrenal medullary cells. Brain Res 1988; 28(454(1â€“2)): 383â€“386. CrossRef
Smiley S.T., Reers M., Mottola-Hartshorn C. et al. Intracellular heterogeneity in mitochondrial membrane potential revealed by a j-aggregate forming lipophilic cation JC-1. Proc. Natl. Acad. Sci. USA 1991; 88: 3671â€“3675. CrossRef
Tze W.J., Tai J., Kardak A. Successful banking of pancreatic endocrine cells for transplantation. Metabolism 1990; 39(7): 719â€“723. CrossRef
Woollacott A.J., Simpson P.B. High throughput fluorescence assays for the measurement of mitochondrial activity in intact human neuroblastoma cells. J Biomol Screen 2001; 6(6): 413â€“420. CrossRef
Zhang W., Wang M., Xie H.Y. et al. Role of reactive oxygen species in mediating hepatic ischemia-reperfusion injury and its therapeutic applications in liver transplantation. Transplant Proc 2007; 39(5): 1332â€“1337. CrossRef PubMed
Yurchuk T.A., Bozhok G.A., Borovoy I.A. et al. Effect of cryopreservation in serum-free and serum-enriched cryoprotective media on formation of j-aggregates in rat adrenal cortex cells. Problems of Cryobiology 2011; 21(4): 408â€“415.
How to Cite
Copyright (c) 2020 Taisiya A. Yurchuk, Galina A. Bozhok, Natalia M. Alabedalkarim, Tatyana P. Bondarenko
This work is licensed under a Creative Commons Attribution 4.0 International License.
Authors who publish with this journal agree to the following terms:
- Authors retain copyright and grant the journal right of first publication with the work simultaneously licensed under a Creative Commons Attribution License that allows others to share the work with an acknowledgement of the work's authorship and initial publication in this journal.
- Authors are able to enter into separate, additional contractual arrangements for the non-exclusive distribution of the journal's published version of the work (e.g., post it to an institutional repository or publish it in a book), with an acknowledgement of its initial publication in this journal.
- Authors are permitted and encouraged to post their work online (e.g., in institutional repositories or on their website) prior to and during the submission process, as it can lead to productive exchanges, as well as earlier and greater citation of published work (See The Effect of Open Access).