1. Quantitative assessment of ischemic tissue damage in ovarian cortical tissue with or without antioxidant (ascorbic acid) treatment 
S.Samuel Kim, M.D., Hyun Won Yang, Ph.D., Hee Gyoo Kang, Ph.D., Hang Heun Lee, M.Sc., Hoi Chang Lee, M.Sc., Duck Sung Ko, M.Sc., Roger G. Gosden, Ph.D. 
Fertility and Sterility 
Volume 82, Issue 3, Pages (September 2004)
DOI: /j.fertnstert
Copyright © 2004 American Society for Reproductive Medicine Terms and Conditions

2. FIGURE 1
Comparison of ischemic tissue damage at two different transportation temperatures. There was no significant difference in oxygen metabolism when specimens were transported in 2 hours either on ice or at room temperature. When comparing oxygen consumption rates between each incubation period (ischemia time), a significant decrease after 24 hours of incubation was noticed (*P<.001).
Fertility and Sterility  , DOI: ( /j.fertnstert )
Copyright © 2004 American Society for Reproductive Medicine Terms and Conditions

3. FIGURE 2
Effect of ascorbic acid on apoptosis of primordial follicles in cultured bovine ovarian tissue. Apoptotic primordial follicles detected by TUNEL staining were counted as well as the total number of primordial follicles to calculate the rate of apoptosis. The number of apoptotic follicles increased in a time-dependent manner (a significant difference with 24 and 48 hours of incubation compared to 0 hours [a, b, c, d; P<.05 by ANOVA]) in both fresh (A) and frozen/thawed (B) ovarian tissue. Ascorbic acid treatment (50 μg/mL) did not improve follicular survival in either fresh (A) or frozen/thawed (B) ovarian tissue.
Fertility and Sterility  , DOI: ( /j.fertnstert )
Copyright © 2004 American Society for Reproductive Medicine Terms and Conditions

4. FIGURE 3
Effect of ascorbic acid on apoptosis of stromal cells in cultured bovine ovarian tissue. The apoptotic rate of stromal cells increased in a time-dependent manner (a significant difference with 24 and 48 hours of incubation compared to 0 hours [a, b, c, d, e, f, g; P<.05 by ANOVA]). Protective effect of ascorbic acid on stromal cell death was clearly seen in fresh (A) and frozen/thawed (B) bovine ovarian tissues cultured for 24 hours with ascorbic acid (*P<.05), but ascorbic acid treatment (50 μg/mL) did not reduce stromal cell death after 48 hours of incubation.
Fertility and Sterility  , DOI: ( /j.fertnstert )
Copyright © 2004 American Society for Reproductive Medicine Terms and Conditions

5. FIGURE 4
Representative sections of bovine ovarian tissue cultured with or without ascorbic acid for 3 and 24 hours and stained by a TUNEL method. At 3 hours after incubation, there was no difference in TUNEL-positive area between the ascorbic acid untreated (a) and treated groups (b). However, specimens incubated for 24 hours without ascorbic acid (c) showed increased TUNEL-positive areas in the cortex compared to those with ascorbic acid treatment (d). A TUNEL-positive primary follicle (e) was seen in fresh ovarian tissue incubated without ascorbic acid. There were intact primordial follicles (f) despite TUNEL-positive surrounding stromal cells, when ascorbic acid was treated (a, b, c, d; magnification ×40: e, f; magnification ×400).
Fertility and Sterility  , DOI: ( /j.fertnstert )
Copyright © 2004 American Society for Reproductive Medicine Terms and Conditions

6. FIGURE 5
A representative autoradiograph showing DNA fragmentation after incubation with (A) or without (N) 50 μg/mL of ascorbic acid for 3, 24, and 48 hours. The protective effect of ascorbic acid was clearly seen in lane 24A (less prominent DNA laddering) compared to lane 24N in both fresh (A) and frozen/thawed (B) specimens.
Fertility and Sterility  , DOI: ( /j.fertnstert )
Copyright © 2004 American Society for Reproductive Medicine Terms and Conditions