Table 7 Summary of vitrification protocols.

From: Embryo morphokinetics derived from fresh and vitrified bovine oocytes predict blastocyst development and nuclear abnormalities

Solution

VCR-H

VCR-L

Composition

Time

Composition

Time

Equilibration

BS10 + 3% EG

12 min

BS20 + 7.5% EG + 7.5% DMSO

12 min

Vitrification

BS10 + 30% EG + 1.0 M sucrose

30–40 s

BS20 + 15% EG + 15% DMSO + 0.5 M sucrose

30–40 s

Warming 1

BS10 + 0.5 M sucrose

1 min

BS20 + 1 M sucrose

1 min

Warming 2

BS10 + 0.25 M sucrose

3 min

BS20 + 0.5 M sucrose

3 min

Warming 3

BS10 + 0.125 M sucrose

3 min

BS20 + 0.25 M sucrose

5 min

Warming 4

BS10 + 0.0625 M sucrose

3 min

  
  1. The base solution contained Hepes-TCM-199 Hank’s with 10% (v/v) (BS10) or 20% (v/v) (BS20) foetal bovine serum. For vitrification, oocytes were placed in 500 μL of equilibration solution (ES) for 10–15 min. and in 100 μL of vitrification solution (VS) for 15 s, then the cryo-device was loaded and plunged in liquid nitrogen (total exposure to VS of 40 s). For warming, oocytes were immersed in 4 mL of warming solution, then washed in 500 µL of the different warming solutions depending on the protocol. After the last warming solution, oocytes were washed two times in BS10 or BS20. EG: ethylene glycol; DMSO: dimethyl sulfoxide. M: mol/L. VCR-H: corona radiata oocytes vitrified with a protocol using high concentrations of cryoprotectants (15%) in equilibration solution; VCR-L: corona radiata oocytes vitrified with a protocol using a low cryoprotectant concentration (3%) in equilibration solution.
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