Rabbit Embryo Vitrification Using Cryogenic Straw: An Ultra-Rapid Cooling Technique to Preserve Rabbit Embryos for Long-term Storage

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

1. 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 dragging the minimum volume can be suitable.
2. Vitrify the embryos in a two-step addition procedure:
   1. Place the embryos for 2 minute....

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 magnif.......

Materials

List of materials used in this article

Name	Company	Catalog Number	Comments
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