Rabbit Embryo Vitrification Using Cryogenic Straw: An Ultra-Rapid Cooling Technique to Preserve Rabbit Embryos for Long-term Storage
Published: April 30, 2023
Abstract
Source: Garcia-Dominguez, X. et al. Minimally Invasive Embryo Transfer and Embryo Vitrification at the Optimal Embryo Stage in Rabbit Model. J. Vis. Exp. (2019)
In this video, we describe the procedure for vitrification of harvested rabbit embryos at desired development stage using ethylene glycol and dimethyl sulfoxide as cryoprotectants and cryogenic straw as the carrier system. The vitrified embryos have high survival rate and can be stored for an extended period until use.
Protocol
All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board. 1. Embryo Vitrification Perform all the manipulations at room temperature (around 22 °C) to reduce the vitrification solution toxicity at warmer temperatures. NOTE: Embryos can be moved using 0.1–2 µL automatic pipette in this protocol, but other similar devices to move the embryos d…
Representative Results
Figure 1: Schematization of correctly loaded straw. A) BM refers to the embryo manipulating media employed during vitrification. Embryos must be loaded in vitrification solution. B) Macroscopic appearance of the loaded straw with a magnified detail of the embryo position. This large-volume device allows us to vitrify large number of embryos, unlike minimum volume devices. Further…
Declarações
The authors have nothing to disclose.
Materials
| Bovine Serum Albumin (BSA) | VWR | 332 | |
| Dimethyl Sulfoxide | Sigma Aldrich | W387509 | |
| Dulbecco's phosphate-buffered saline (DPBS) | Sigma Aldrich | D5773 | Without calcium chloride. |
| Ethylene Glycol | Sigma Aldrich | 102466-M | |
| Liquid Nitrogen | Air Liquide | P1505XXX | |
| Plastic Straw 0.25 mL | IMV – technologies | 6431 | |
| Stereomicroscope | Leica | MZ16F | There are cheaper options such as Leica MZ8 or Nikon SMZ-10 or SMZ-2B, to name a few. |
| Straw Plug | IMV – technologies | 6431 |
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Rabbit Embryo Vitrification Using Cryogenic Straw: An Ultra-Rapid Cooling Technique to Preserve Rabbit Embryos for Long-term Storage. J. Vis. Exp. (Pending Publication), e20935, doi: (2023).