Abstract
At present, there are three ways to determine effectively the quality of the cryopreservation procedure using ovarian tissue before the re-implantation treatment: evaluation of follicles after post-thawing xenotransplantation to SCID mouse, in-vitro culture in a large volume of culture medium under constant agitation and culture on embryonic chorio-allantoic membrane within a hen's eggs. The aim of this study was to compare the two methods, culture in vitro and culture on embryonic chorioallantoic membrane (CAM) of cryopreserved human ovarian medulla-contained and medulla-free cortex. Ovarian fragments were divided into small pieces (1.5–2.0×1.0–1.2×0.8–1.5) of two types, cortex with medulla and medulla-free cortex, frozen, thawed and randomly divided into the following four groups. Group 1: medulla-free cortex cultured in vitro for 8 days in large volume of medium with mechanical agitation, Group 2: medulla-containing cortex cultured in vitro, Group 3: medulla-free cortex cultured in CAM-system for 5 days, Group 4: medulla-containing cortex cultured in CAM-system. The efficacy of the tissue culture was evaluated by the development of follicles and by intensiveness of angiogenesis in the tissue (von Willebrand factor and Desmin). For Group 1, 2, 3 and 4, respectively 85%, 85%, 87% and 84% of the follicles were morphologically normal (P>0.1). The immunohistochemical analysis showed that angiogenesis detected by von Willebrand factor was lower in groups 1 and 3 (medulla-free cortex). Neo-vascularisation (by Desmin) was observed only in ovarian tissue of Group 4 (medulla-contained cortex after CAM-culture). It appears that the presence of medulla in ovarian pieces is beneficial for post-thaw development of cryopreserved human ovarian tissue. For medical practice it is recommended for evaluation of post-warming ovarian tissue to use the CAM-system as a valuable alternative to xenotransplantation and for cryopreservation of these tissues to prepare ovarian medulla-contained strips.
Highlights
Cancer is one of the major death causes: in the USA alone a total of 1,596,670 new cancer cases will be reported in 2011, of which the projected number of cancer-related deaths were 571,950
Overall cancer incidence rates in women in the USA have been declining by 0.6% annually since 1998 [1]
The current estimate for 2010 for Germany relates to a total of approximately 204,000 cancer cases in women, and every year in Germany, around 800 girls under age 15 are diagnosed with cancer [3]
Summary
Cancer is one of the major death causes: in the USA alone a total of 1,596,670 new cancer cases will be reported in 2011, of which the projected number of cancer-related deaths were 571,950. Overall cancer incidence rates in women in the USA have been declining by 0.6% annually since 1998 [1]. The overall incidence rate for cancer in children aged 14 years and younger increased by 0.6% per year between 1998 and 2007 [2]. The current estimate for 2010 for Germany relates to a total of approximately 204,000 cancer cases in women, and every year in Germany, around 800 girls under age 15 are diagnosed with cancer [3]. The death rate has been decreasing by 1.0% per year
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