Logo Medical Science Monitor Basic Research

Call: 1.631.470.9640
Mon-Fri 10 am - 2 pm EST

Contact Us

Logo Medical Science Monitor Basic Research Logo Medical Science Monitor Basic Research Logo Medical Science Monitor Basic Research

04 October 2013

Cytoskeletal alterations in different developmental stages of in vivo cryopreserved preimplantation murine embryos

Razif DasimanABCDEF, Nor-Shahida Abdul RahmanBCD, Salina OthmanBCD, Mohd-Fazirul MustafaCD, Norhazlin JusohCD, Wan-Hafizah W. JusofCD, Mohd Hamim RajikinADE, Gabriele Ruth Anisah FroemmingADE, Nor-Ashikin Mohamed Noor KhanADEFG

DOI: 10.12659/MSMBR.884019

Med Sci Monit Basic Res 2013; 19:258-266

Abstract

BACKGROUND: This study aimed to investigate the effects of vitrification and slow freezing on actin, tubulin, and nuclei of in vivo preimplantation murine embryos at various developmental stages using a Confocal Laser Scanning Microscope (CLSM).

MATERIAL AND METHODS: Fifty female mice, aged 4–6 weeks, were used in this study. Animals were superovulated, cohabitated overnight, and sacrificed. Fallopian tubes were excised and flushed. Embryos at the 2-cell stage were collected and cultured to obtain 4- and 8-cell stages before being cryopreserved using vitrification and slow freezing. Fixed embryos were stained with fluorescence-labelled antibodies against actin and tubulin, as well as DAPI for staining the nucleus. Labelled embryos were scanned using CLSM and images were analyzed with Q-Win software V3.

RESULTS: The fluorescence intensity of both vitrified and slow-frozen embryos was significantly lower for tubulin, actin, and nucleus as compared to non-cryopreserved embryos (p<0.001). Intensities of tubulin, actin, and nucleus in each stage were also decreased in vitrified and slow-frozen groups as compared to non-cryopreserved embryos.

CONCLUSIONS: Cryopreservation of mouse embryos by slow freezing had a more detrimental effect on the actin, tubulin, and nucleus structure of the embryos compared to vitrification. Vitrification is therefore superior to slow freezing in terms of embryonic cryotolerance.

Keywords: Blastocyst - metabolism, Cell Nucleus - metabolism, Cryopreservation, Cytoskeleton - metabolism, Embryonic Development, Fluorescence, Mice, Inbred ICR, Microscopy, Confocal, Tubulin - metabolism

Add Comment 0 Comments

729 4

Most Viewed Current Articles

05 Jan 2021 : Review article  

A Southeast Asian Perspective on the COVID-19 Pandemic: Hemoglobin E (HbE)-Trait Confers Resistance Against...

DOI :10.12659/MSMBR.929207

Med Sci Monit Basic Res 2021; 27:e929207

05 May 2022 : Laboratory Research  

Calcitriol Inhibits Proliferation and Potentially Induces Apoptosis in B16-F10 Cells

DOI :10.12659/MSMBR.935139

Med Sci Monit Basic Res 2022; 28:e935139

09 Jun 2021 : Laboratory Research  

Vitamin D Inhibits Lipopolysaccharide (LPS)-Induced Inflammation in A549 Cells by Downregulating Inflammato...

DOI :10.12659/MSMBR.931481

Med Sci Monit Basic Res 2021; 27:e931481

07 Jul 2022 : Laboratory Research  

Cytotoxicity, Apoptosis, Migration Inhibition, and Autophagy-Induced by Crude Ricin from Ricinus communis S...

DOI :10.12659/MSMBR.936683

Med Sci Monit Basic Res 2022; 28:e936683

Your Privacy

We use cookies to ensure the functionality of our website, to personalize content and advertising, to provide social media features, and to analyze our traffic. If you allow us to do so, we also inform our social media, advertising and analysis partners about your use of our website, You can decise for yourself which categories you you want to deny or allow. Please note that based on your settings not all functionalities of the site are available. View our privacy policy.

Medical Science Monitor Basic Research eISSN: 2325-4416
Medical Science Monitor Basic Research eISSN: 2325-4416