Table 1.
Derivative solute concentrations in various dilutions of the EAFS10/10 vitrification solution.
Table 2.
Mass compositions of various dilutions of EAFS10/10 vitrification solution.
Table 3.
Protocol of each cooling procedure.
Table 4.
Protocol of each warming procedure.
Figure 1.
Survival of mouse oocytes as a function of the dilution of EAFS and the cooling and warming rate to and from −196°C.
The dilutions were 1.0×, 0.875×, 0.75×, or 0.5×. The cooling rates ranged from 37°C/min to 69,250°C/min; the warming rates (°C/min) were 117,500 (Δ). 2,950 (•). and 2,170 (▪). The protocols are given in Tables 3 and 4. Survivals are based on morphological appearance and osmotic behavior after warming and after 1–2 hr incubation in M16 medium.
Table 5.
Survival based on morphological normality and osmotic responsiveness of mouse oocytes suspended in various dilution of EAFS10/10, and cooled and warmed at indicated rates in straws or on Cryotops.
Figure 2.
Survival of mouse oocytes vs. the relative concentration of the EAFS vitrification solution in which they were suspended.
The relative concentrations ranged from 0.33× to 1×. The absolute concentrations of the individual solutes are given in Table 1 and 2. The suspensions were cooled to −196°C at rates ranging from 522 to 69,250°C/min and subsequently warmed at five different rates. The oocytes in the curves delineated by the open circles and triangles were warmed on Cryotops at the highest attainable rate of 117,500°C/min. The cells in the curves delineated by the closed symbols were warmed at lower rates between 610 and 2,950°C/min. CR and WR stand for cooling rate and warming rate. Symbol CR (°C/min) WR (°C/min) Δ 69,250 117,500 ○ 880 117,500 ▴ 69,250 610 ▪ 522 2,950 ♦ 522 2,170 Survivals are based on morphological appearance and osmotic behavior after 1–2 hr incubation in M16 medium.
Table 6.
Morphological and functional survival of mouse oocytes suspended in various dilution of EAFS10/10, cooled at indicated rates on Cryotops, and warmed at 117,500°C/min.