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., SMETANSKA, I.: Effects of elicitors and high hydrostatic pressure on secondary metabolism of Vitis vinifera suspension culture. Process Biochem., 46, 2011, 1411-1416. DELGADO ADÁMEZ, J., GAMERO SAMINO, E., VALDÉS SÁNCHEZ, E., GONZÁLEZ-GÓMEZ, D.: In vitro estimation of the antibacterial activity and antioxidant capacity of aqueous extracts from grape-seeds (Vitis vinifera L.). Food Control, 24, 2012, 136-141. DÖRNENBURG, H., KNORR, D.: Strategies for the improvement of secondary metabolite production in plant cell cultures. Enzyme Microb. Technol., 17, 1995, 674

time congruent with crew duties. However, the rapid and consistent defrosting of cell cultures is challenging in a microgravitational environment. Frozen cells on Earth (typically in a cryovial) may be rapidly thawed by agitation in a 37°C water bath. However, the use of an open water bath is not possible on the ISS. We here describe the development of a contained system that allows the rapid thawing of adherent cell cultures. COMPARATIVE DEFROST METHODS In developing an on-orbit defrost system, we examined alternative methods to rapidly defrost an enclosed large

1 Introduction Experimentation using in vitro cell culture serves as a foundation of biomedical research. Immortalized cell lines and primary cells are cultured and maintained in cell culture media; a specially formulated mixture of metabolites that supports cellular growth and proliferation. Widely used basal medias (e.g. DMEM, RPMI, MEM) are manufactured with the intent of minimizing variability between culture techniques ( Asayama, 2017 ). These basal media are further supplemented with various compounds to fit the needs of the specific cell line or type

DMEM supplemented with 10% fetal calf serum (FCS; Merck, Darmstadt, Germany) and 10 U/ mL penicillin G, 10 mg/mL streptomycin, and 25 μg/ mL amphotericin B. Cell viability was 90 to 95% as determined by trypan blue staining (Merck, Darmstadt, Germany). The cells were cultured at 37 °C in a humidified atmosphere of 5% CO2. Once the cell cultures attained 70–80% confluency, they were passaged by washing with PBS, digested with 0.025% Trypsin/EDTA (Cascade Biologics, Portland, USA), neutralized by a 0.0125% trypsin inhibitor (Cascade Biologics, Portland, USA

( Bubalus bubalis ) embryos. Reprod. Domest. Anim., 53: 986–996. Aoued H.S., Singh M. (2015). Recovery of fibroblast-like cells after 160 days of postmortem storage of goat skin tissues in refrigerated media. J. Veterinar. Sci. Technol., 6: 236. Balin A.K., Fisher A.J., Anzelone M., Leong I., Allen R.G. (2002). Effects of establishing cell cultures and cell culture conditions on the proliferative life span of human fibroblasts isolated from different tissues and donors of different ages. Exp. Cell Res., 274: 275–287. Barros F.R.de,Goissis M.D., Caetano H.V., Paula-Lopes F

References Ahne W., Negele R.D. 1985 - Studies on the transmission of infectious pancreatic necrosis virus via eyed eggs and sexual products of salmonid fish - In: Fish and Shellfish Pathology (Ed.) A.E. Ellis, Academic Press, London: 262-270. Alonso M.C., Cano I., Castro D., Perez-Prieto S.I., Borrego J.J. 2004 - Development of an in situ hybridization procedure for the detection of sole aquabirnavirus in infected fish cell cultures - J. Virol. Methods 116:133-138. Ball H.J., Munro A.L.S., Ellia A., Elson K.G.R., Hodgkiss W., Farlane L.S. 1971 - Infectious

REFERENCES Astashkina, A., Mann, B., Grainger, D. W. (2012). A critical evaluation of in vitro cell culture models for high-throughput drug screening and toxicity. Pharmacol. Ther ., 134 , 82–106. Azadi, M., Jamali, T., Kianmehr, Z., Kavoosi, G., Ardestani, S. K. (2020). In-vitro (2D and 3D cultures) and in-vivo cytotoxic properties of Zataria multiflora essential oil (ZEO) emulsion in breast and cervical cancer cells along with the investigation of immunomodulatory potential. J. Ethnopharmacol ., 257 , 112865. Baker, B. M., Chen, C. S. (2012

References Andersen CY, Jørgensen N (1995). Improvement of sperm motility by the addition of progesterone to the Percoll medium during sperm purifi cation. Hum Reprod 10: 3183-3185. Bates MK (2012). Culturing cells under hypoxic conditions for biologically relevant results. Am Lab. Carreau A, Hafny-Rahbi BE, Matejuk A, Grillon C, Kieda C (2011). Why is the partial oxygen pressure of human tissues a crucial parameter? Small molecules and hypoxia. J Cell Mol Med 15: 1239-1253. Gille JJP, Joenje H (1992). Cell culture models for oxidative stress: Superoxide and

print] Butler M. (2004). Animal cell culture and technology. 2a Ed. London, England: Taylor & Francis. Česen MH, Pegan K, Spes A, Turk B. (2012). Lysosomal pathways to cell death and their therapeutic applications. Exp Cell Res, 318, 1245-51. Fotakis G, Timbrell JA. (2006). In vitro cytotoxicity assays: comparison of LDH, neutral red, MTT and protein assay in hepatoma cell lines following exposure to cadmium chloride. Toxicol Lett, 160, 171-7. Freshney RI. (2005). Culture of animal cells: a manual of basic technique. 5a. Ed. New Jersey, U.S.: Wiley

, sensitivity, and detection mechanism, have to be considered ( 4 ). The mechanism of cell cytotoxicity demonstrated by an active substance is for clarification in these circumstances, and for this purpose a cell culture system is required for quantitative assay of malignant transformation by the substance. BALB/3T3 mouse fibroblast cells were used in our study to examine the cytotoxic effect of inosine pranobex, since the BALB/3T3 system was shown to be sensitive to a wide diversity of potential carcinogens or procarcinogens ( 14 ). Human hepatoma HepG2 cells display many