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Microscopic Assessment of Dead Cell Ratio in Cryopreserved Chicken Primordial Germ Cells

Part of: Micrographia Collection

Published online by Cambridge University Press:  18 September 2019

Andrea Svoradová Open the ORCID record for Andrea Svoradová [Opens in a new window] ,
Alexander Makarevich ,
Jaromír Vašíček ,
Lucia Olexiková ,
Sasa Dragin  and
Peter Chrenek
Andrea Svoradová*
Affiliation:
Faculty of Natural Sciences, Constantine the Philosopher University, Trieda A. Hlinku 1, 949 79 Nitra, Slovakia
Alexander Makarevich
Affiliation:
Research Institute for Animal Production in Nitra, National Agricultural and Food Centre, Hlohovecká 2, 951 41 Lužianky, Slovakia
Jaromír Vašíček
Affiliation:
Research Institute for Animal Production in Nitra, National Agricultural and Food Centre, Hlohovecká 2, 951 41 Lužianky, Slovakia Faculty of Biotechnology and Food Science, Slovak University of Agriculture, Trieda A. Hlinku 2, 949 76 Nitra, Slovakia
Lucia Olexiková
Affiliation:
Research Institute for Animal Production in Nitra, National Agricultural and Food Centre, Hlohovecká 2, 951 41 Lužianky, Slovakia
Sasa Dragin
Affiliation:
Faculty of Agriculture, University of Novi Sad, Trg Dositeja Obradovića 8, Novi Sad, Serbia
Peter Chrenek
Affiliation:
Research Institute for Animal Production in Nitra, National Agricultural and Food Centre, Hlohovecká 2, 951 41 Lužianky, Slovakia Faculty of Biotechnology and Food Science, Slovak University of Agriculture, Trieda A. Hlinku 2, 949 76 Nitra, Slovakia Department of Animal Biochemistry and Biotechnology, University of Science and Technology, Al. prof. S. Kaliskiego 7, 85-796 Bydgoszcz, Poland
*
*Author for correspondence: Andrea Svoradová, E-mail: svoradovaandrea1@gmail.com
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Abstract

This study aimed to compare three methods of cell death assessment [trypan blue exclusion (TBE), propidium iodide viability assay (PIVA), and transmission electron microscopy] to evaluate fresh and frozen–thawed chicken primordial germ cells (PGCs). For this study, chicken PGCs were collected from ROSS 908 and Oravka breed hens, cryopreserved-thawed according to the protocol, and submitted for different cell death assessments. We observed significant differences between TBE and PIVA techniques in the detectable proportion of dead cells in fresh (14.14 ± 1.27 versus 7.16 ± 1.02%, respectively) and frozen–thawed (44.00 ± 2.11 versus 33.33 ± 1.67%, respectively) samples of the Oravka breed. Moreover, significant differences (p < 0.05) between TBE and PIVA techniques in the detectable proportion of dead cells in fresh (9.20 ± 0.60 versus 5.37 ± 0.51%) samples of ROSS 908 breed were recorded. Differences may be due to methodological, sensitivity, and toxicity features of each technique tested, where TB stains cell cytoplasm of dead cells and PI penetrates and intercalates into DNA of dead cells. Therefore, we suggest using a more precise and sensitive PIVA for viability evaluation of PGCs. Further research is needed to apply various fluorochromes for more detailed cell viability evaluation.

Keywords

cryopreservationelectron microscopyprimordial germ cellsviability
Type
Micrographia
Information
Microscopy and Microanalysis , Volume 25 , Issue 5 , October 2019 , pp. 1257 - 1262
Copyright
Copyright © Microscopy Society of America 2019 

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References

Altmanm, SA, Randers, L & Rao, G (1993). Comparison of trypan blue dye exclusion and fluorometric assays for mammalian cell viability determinations. Biotechnol Prog 9(6), 671–674.Google Scholar
Baskić, D, Popović, S, Ristić, P & Arsenijević, NN (2006). Analysis of cycloheximide-induced apoptosis in human leukocytes: fluorescence microscopy using annexin V/propidium iodide versus acridin orange/ethidium bromide. Cell Biol Int 30(11), 924–932.Google Scholar
Chojnacka-Puchta, L, Sawicka, D, Lakota, P, Plucienniczak, G, Bednarczyk, M & Plucienniczak, A (2015). Obtaining chicken primordial germ cells used for gene transfer: In vitro and in vivo results. J Appl Genet 56(4), 493–504.Google Scholar
Elmore, S (2007). Apoptosis: A review of programmed cell death. Toxicol Pathol 35(4), 495–516.Google Scholar
Glover, JD & McGrew, MJ (2012). Primordial germ cells technologies for avian germplasm cryopreservation and investigating germ cell development. J Poultry Sci 49, 155–162.Google Scholar
Idziorek, T, Estaquier, J, De Bels, F & Ameisen, JC (1995). YOPRO-1 permits cytofluorometric analysis of programmed cell death (apoptosis) without interfering with cell viability. J Immunol Methods 185, 249–258.Google Scholar
Kohara, Y, Kanai, Y & Tajima, A (2008). Cryopreservation of gonadal germ cells (GGCs) from the domestic chicken using vitrification. J Poultry Sci 45, 57–61.Google Scholar
Kostaman, T, Yusuf, TL, Fachrudin, M & Setiadi, MA (2018). Effectiveness of DMSO concentration on recovery rate and viability of primordial germ cell of Gaok chicken. Indones J Anim Vet Adv 22(1), 30–37.Google Scholar
Kummrow, A, Frankowski, M, Bock, N, Werner, C, Dziekan, T & Neukammer, J (2013). Quantitative assessment of cell viability based on flow cytometry and microscopy. Cytometry Part A 83(2), 197–204.Google Scholar
Li, BC, Chen, GH, Qin, J, Wu, XS, Wu, SL & Cai, ZT (2005). Suitable stages for isolation and culture PGCs from chicken embryos. Int J Poult Sci 4(11), 885–890.Google Scholar
Lichtman, JW & Conchello, JA (2005). Fluorescence microscopy. Nat Methods 2(12), 910–919.Google Scholar
Mascotti, K, McCullough, J & Burger, SR (2000). HPC viability measurement: Trypan blue versus acridine orange and propidium iodide. Transfusion 40(6), 693–696.Google Scholar
Minematsu, T, Tajima, A & Kanai, Y (2004). The migratory ability of gonadal germ cells in the domestic chicken. J Poultry Sci 41, 178–185.Google Scholar
Moore, DT, Purdy, PH & Blackburn, HD (2006). A method for cryopreserving chicken primordial germ cells. Poult Sci 85(10), 1784–1790.Google Scholar
Naito, M (2003). Cryopreservation of avian germline cells and subsequent production of viable offspring. J Poultry Sci 40, 1–12.Google Scholar
Nakajima, Y, Minematsu, T, Naito, M & Tajima, A (2011). A new method for isolation viable gonadal germ cells from 7-day-old chick embrios. J Poultry Sci 48, 106–111.Google Scholar
Pavlova, I, Williams, M, El-Naggar, A, Richards-Kortum, R & Gillenwater, A (2008). Understanding the biological basis of autofluorescence imaging for oral cancer detection: High-resolution fluorescence microscopy in viable tissue. Clin Cancer Res 14(8), 2396–2404.Google Scholar
Qian, C, Zhau, Z, Han, H, Zhao, C, Jin, X, Zhao, H, Zhang, Y, Chen, W, Yang, N & Li, Z (2010). Influence of microgravity on the concentration of circulating primordial germ cells in silky chicken offspring. J Poultry Sci 47, 65–70.Google Scholar
Reissis, Y, García-Gareta, E, Korda, M, Blunn, GW & Hua, J (2013). The effect of temperature on the viability of human mesenchymal stem cells. Stem Cell Res Ther 4(6), 139.Google Scholar
Riccardi, C & Nicoletti, I (2006). Analysis of apoptosis by propidium iodide staining and flow cytometry. Nat Protoc 1(3), 1458.Google Scholar
Sawicka, D, Brzezińska, J & Bednarczyk, M (2011). Cryoconservation of embryonic cells and gametes as a poultry biodiversity preservation method. Folia Biol 59, 1–5.Google Scholar
Sawicka, D, Chojnacka-Puchta, L, Zielinski, M, Plucienniczak, G, Plucienniczak, A & Bednarczyk, M (2015). Flow cytometric analysis of apoptosis in cryoconserved chicken primordial germ cells. Cell Mol Biol Lett 20(1), 143–159.Google Scholar
Setioko, AR, Tagami, T, Tase, H, Nakamura, Y, Takeda, K & Nirasawa, K (2007). Cryopreservation of primordial germ cells (PGCs) from White Leghorn embryos using commercial cryoprotectants. J Poultry Sci 44(1), 73–77.Google Scholar
van de Lavoir, MC, Diamond, JH, Leighton, PA, Mather-Love, C, Heyer, BS, Bradshaw, R, Kerchner, A, Hooi, LT, Gessaro, TM, Swanberg, SE, Delany, ME & Etches, RJ (2006). Germline transmission of genetically modified primordial germ cells. Nature 441, 766–769.Google Scholar
Zhang, Y, Chen, X, Gueydan, C & Han, J (2018). Plasma membrane changes during programmed cell deaths. Cell Res 28(1), 9.Google Scholar