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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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