1. Histologic and ultrastructural features of cryopreserved ovine ovarian tissue: deleterious effect of 1,2-propanediol applying different thawing protocols 
Irma C. Oskam, Ph.D., Babak A. Asadi, M.Sc., Regiane R. Santos, Ph.D. 
Fertility and Sterility 
Volume 93, Issue 8, Pages (May 2010)
DOI: /j.fertnstert
Copyright © 2010 American Society for Reproductive Medicine Terms and Conditions

2. Figure 1
(A) Total additive stroma scores (i.e., normal morphology, pyknotic cells, and edema) recorded in control and treated ovarian tissue. (B) Mean percentages (± SE) of morphologically normal preantral follicles after freezing-thawing of ovine ovarian tissue in the presence of ethylene glycol (EG) or 1,2-propanediol (PROH); thawing was performed at 37°C for EG and 4°, 30°, or 37°C for PROH. (C) Panel with light microscopic images of control (CIa–CIc) and ovine ovarian tissue fragments cryopreserved in presence of PROH (CIIa–CIIc) and EG (CIIIa–CIIIc). Healthy and preserved stroma and follicles are observed in control (CIa and CIb) and EG (CIIIa and CIIIb) images, whereas image CIIa and CIIb (PROH 30°C) show degenerating preantral follicles and affected stroma. Affected stroma (i.e., reduced density of collagen [stained in blue] and fibrin [stained in red]) after cryopreservation in presence of PROH could be observed after Acid Fuchsin Orange G (AFOG) staining CIIc. (D) Transmission electron microscopy revealed a normal primordial follicle frozen and thawed in presence of EG. The oocyte (Oo) is rather well preserved, although granulosa cells (GC) present a polarization of cytomicroorganeles (DIa); see details at DIb. After freezing-thawing in presence of PROH, in general the ovarian fragment showed an irreversibly damaged primordial follicle with a shrunken nucleus (Nu) and cytoplasm presenting multiple vacuoles (MV), and the surrounding stroma is compromised (DIIa). In addition, GCs are swollen, do not present contact with the oocyte, and reveal cytoplasm either empty or with a low density of organelles (DIIb). ∗Differs significantly from control (P<.05); a–c, lowercase letters in A indicate significant differences among treatments (P<.05). Thawing at 30°C.
Fertility and Sterility  , DOI: ( /j.fertnstert )
Copyright © 2010 American Society for Reproductive Medicine Terms and Conditions