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Modified MicroSecure Vitrification: A Safe, Simple and Highly Effective Cryopreservation Procedure for Human Blastocysts
Published on: March 2, 2017
Successful cryopreservation of expanded equine blastocysts
Y H Choi1, I C Velez, F L Riera
1College of Veterinary Medicine & Biomedical Sciences, Texas A&M University, College Station, Texas 77843, USA.
Theriogenology
|April 5, 2011
Summary
Cryopreservation of equine blastocysts is challenging. This study shows that inducing blastocyst collapse via biopsy and vitrifying in fine pipettes significantly improves survival rates for expanded equine embryos.
Area of Science:
- Reproductive Biology
- Cryobiology
- Equine Science
Background:
- Effective cryopreservation of expanded equine blastocysts is hindered by factors like large blastocoele fluid volume and the embryonic capsule.
- Previous research indicated successful viability of equine embryos post-trophoblast biopsy, which naturally caused blastocyst collapse.
Purpose of the Study:
- To investigate the impact of trophoblast biopsy, capsule breach, and resultant blastocyst collapse on the survival of expanded equine blastocysts during vitrification.
- To optimize vitrification protocols for equine blastocysts, focusing on techniques that promote blastocoele fluid loss and collapse.
Main Methods:
- Vitrification of small (< 300 μm) and expanded (300-730 μm) equine blastocysts using dimethylsulfoxide (DM) or ethylene glycol (EG) based media in fine-diameter pipette tips.
- Comparison of warming protocols, including sucrose (EG/s), and evaluation of different biopsy techniques (standard vs. central aspiration) to remove blastocoele fluid.
- Assessment of embryo survival and pregnancy rates (via embryonic heartbeat detection) following warming and embryo transfer.
Main Results:
- Vitrification using DM and EG/s media resulted in pregnancies for non-biopsied embryos (3/12 and 3/6, respectively).
- Biopsied and vitrified expanded blastocysts showed improved pregnancy rates with EG/s (46%) compared to DM (13%).
- Techniques involving blastocoele fluid aspiration prior to vitrification, particularly with EG/s, led to higher pregnancy rates, reaching 71% (5/7) in the final optimized protocol.
Conclusions:
- Inducing blastocoele collapse through biopsy and subsequent vitrification in fine-diameter pipettes is crucial for successful cryopreservation of expanded equine blastocysts.
- The optimized method involving peripheral biopsy, blastocoele fluid aspiration, and vitrification in EG/s medium significantly enhances the viability and pregnancy rates of equine blastocysts.
