Edorium Journal of

Anatomy and Embryology

 
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Original Article
 
Using the balance between proliferation and apoptosis to assess the cryopreservation and thawing protocol in mouse 4-cell embryos
Mostafa M. El-Naggar1, Hassan Nasrat2, Hassan Jamal2, Samar Al-Saggaf3, Mohamed H. Badawod3
1Professor, Department of Anatomy, Faculty of Medicine, Jazan University, Jazan, Saudi Arabia.
2Professor, Department of Obstetrics and Gynecology, Faculty of Medicine, King Abdulaziz University, Jeddah, Saudi Arabia.
3Professor, Department of Anatomy, Faculty of Medicine, King Abdulaziz University, Jeddah, Saudi Arabia.

Article ID: 100003A04ME2015
doi:10.5348/A04-2015-3-OA-2

Address correspondence to:
Dr. Mostafa M. El-Naggar
Faculty of Medicine, Jazan University
P.O. Box 114
Jazan
Kingdom of Saudi Arabia
Phone: + (966) 506570529

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How to cite this article
El-Naggar MM, Nasrat H, Jamal H, Al-Saggaf S, Badawod MH. Using the balance between proliferation and apoptosis to assess the cryopreservation and thawing protocol in mouse 4-cell embryos. Edorium J Anat Embryo 2015;2:6–13.


Abstract
Aims: The criteria used to assess the optimal conditions for cryopreservation of the embryos in the in vitro fertilization (IVF) protocols are still a matter of discussion. This study aimed at evaluating the use of cell proliferation and apoptosis to assess the optimal conditions for cryopreservation/thawing of the 4-cell embryos.
Methods: Fertilized ova were collected from mated female MF1 mice 24 hours after hCG injection. They were cultured in KSOM medium and kept in CO2 incubator at 37°C and 5% CO2 up to the stage of the 4-cells. Two methods of cryopreservation were used; the step-rate and the ultra-rapid vitrification. Slow and fast thawing was done. Slides were prepared from samples of the embryos, and stained immunohistochemically for proliferative and apoptotic cells. The proliferative capacity was measured by labeling with bromodeoxyuridine (BrdU) and the apoptotic ability was measured with TUNEL technique.
Results: Vitrification with fast thawing of the 4-cell embryos gave better morphology, higher proliferative capacity, and lower apoptotic ability. Following step-rate cryopreservation with slow or fast thawing, cell labeling index for BrdU was 0% and 17%, respectively and was 66% and 83%, respectively following vitrification. The incidence of apoptosis following step rate cryopreservation with slow or fast thawing was 96% and 89%, respectively and was 42% and 13%, respectively following vitrification.
Conclusion: It is concluded that cell proliferation and apoptosis could be used to assess the cryopreservation/thawing protocol for early embryos.

Keywords: Apoptosis, Cryopreservation, Embryos, Mouse, Proliferation, Thawing

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Author Contributions:
Mostafa M. El-Naggar – Substantial contributions to conception and design, Acquisition of data, Analysis and interpretation of data, Drafting the article, Revising it critically for important intellectual content, Final approval of the version to be published
Hassan Nasrat – Substantial contributions to conception and design, Acquisition of data, Analysis and interpretation of data, Drafting the article, Revising it critically for important intellectual content, Final approval of the version to be published
Hassan Jamal – Substantial contributions to conception and design, Acquisition of data, Analysis and interpretation of data, Drafting the article, Revising it critically for important intellectual content, Final approval of the version to be published
Samar Al-Saggaf – Substantial contributions to conception and design, Acquisition of data, Analysis and interpretation of data, Drafting the article, Revising it critically for important intellectual content, Final approval of the version to be published
Mohamed H. Badawod – Substantial contributions to conception and design, Acquisition of data, Analysis and interpretation of data, Drafting the article, Revising it critically for important intellectual content, Final approval of the version to be published
Guarantor of submission
The corresponding author is the guarantor of submission.
Source of support
None
Conflict of interest
Authors declare no conflict of interest.
Copyright
© 2015 Mostafa M. El-Naggar et al. This article is distributed under the terms of Creative Commons Attribution License which permits unrestricted use, distribution and reproduction in any medium provided the original author(s) and original publisher are properly credited. Please see the copyright policy on the journal website for more information.



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