Cryopreservation of fluorescence activated cell sorted boar spermatozoa based on extracellular ubiquitination
The result's identifiers
Result code in IS VaVaI
<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F00216208%3A11310%2F16%3A10329732" target="_blank" >RIV/00216208:11310/16:10329732 - isvavai.cz</a>
Result on the web
<a href="http://dx.doi.org/10.17221/27/2015-CJAS" target="_blank" >http://dx.doi.org/10.17221/27/2015-CJAS</a>
DOI - Digital Object Identifier
<a href="http://dx.doi.org/10.17221/27/2015-CJAS" target="_blank" >10.17221/27/2015-CJAS</a>
Alternative languages
Result language
angličtina
Original language name
Cryopreservation of fluorescence activated cell sorted boar spermatozoa based on extracellular ubiquitination
Original language description
The present study is focused on the methodology of fluorescence activated cell sorting (FACS) of spermatozoa stained by the antibody against extracellular surface marker ubiquitin (eUb) and subsequent protocol for their long term storage in liquid nitrogen (LN). High level of spermatozoa surface ubiquitination has been previously discussed as a negative quality marker. From a general point of view, any other outer membrane antigen would be compatible with our approach. Regarding our experimental design we found that only those insemination doses with at least 40% of motile spermatozoa after freezing and thawing (F/T) in the egg-yolk medium with lactose are suitable for the subsequent antibody staining and FACS. The sorting rate was sufficient for the preparation of up to 20 spermatozoa aliquots for intracytoplasmic sperm injections (ICSI). Two significantly different groups with good freezability were prepared and stored in LN (0.73% contamination of spermatozoa with high eUb level in non-ubiquitinated group and reversely 6.65% spermatozoa without eUb in highly ubiquitinated group). Sperm viability after FACS varied from 11 to 28% regardless of the used media (P = 0.15). Required viability of F/T sorted spermatozoa was obtained by using Solusem (R) extender as a load and collection medium. In this case 12% of viable spermatozoa with progressive motility in low eUb level group and 7% in high eUb level group (P < 0.05) were detected. Our approach allows obtaining sufficient number of viable spermatozoa for subsequent artificial fertilization by ICSI. This procedure could be used for a wide variety of spermatozoa sorting based on different surface markers.
Czech name
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Czech description
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Classification
Type
J<sub>x</sub> - Unclassified - Peer-reviewed scientific article (Jimp, Jsc and Jost)
CEP classification
EB - Genetics and molecular biology
OECD FORD branch
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Result continuities
Project
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Continuities
S - Specificky vyzkum na vysokych skolach<br>I - Institucionalni podpora na dlouhodoby koncepcni rozvoj vyzkumne organizace
Others
Publication year
2016
Confidentiality
S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů
Data specific for result type
Name of the periodical
Czech Journal of Animal Science
ISSN
1212-1819
e-ISSN
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Volume of the periodical
61
Issue of the periodical within the volume
7
Country of publishing house
CZ - CZECH REPUBLIC
Number of pages
7
Pages from-to
310-316
UT code for WoS article
000382555600002
EID of the result in the Scopus database
2-s2.0-84979911715