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Comparison of quartz vials with polypropylene vials for rapid cryopreservation of human ovarian tissue

View Article: PubMed Central - PubMed

ABSTRACT

Background: Because higher survival of follicles during the freezing/thawing procedure improves the quality of cryopreserved tissue reimplanted after oncological therapies, defining an optimal method for human ovarian tissue cryopreservation remains a major issue in this field. One option to improve the cryopreservation procedure is to use better materials, i.e., vials with better conductivity. The aim of this study was to compare polypropylene (PP) with quartz vials. Between September 2012 and January 2013, eight patients were recruited. The ovarian cortex was cut into 3 slices, assigned randomly to a fresh and a cryopreserved group in PP (method B) or quartz vials (method C). Histological and immunohistochemical (IHC) analysis were used. For IHC three antibodies were analyzed: Ki67 (proliferation index), Bcl2 (anti apoptotic index) and Hsp70 (stress index).

Results: The majority of GCs showed positive staining for Bcl2 in both cryopreservation device, with higher expression in group C than in group B. Oocytes and their nuclei showed intense positive staining for ki67 in both methods B and C, and also a patch positive stromal cells staining for Ki67. Expression of hsp70 was not increased after cryopreservation.

Conclusions: Cryopreservation using quartz vials led to larger numbers of good follicles while maintaining consistent preservation for stromal cells and vessels.

No MeSH data available.


Light microscopy images of cortical human ovarian tissue stained with haematoxylin-eosin from fresh (control) tissue (a, d, g), cryopreserved in PP device (b, e, h) and cryopreserved in quartz device (c, f, i). Stromal cells in cryopreserved tissue using PP (b) or quartz (c) vial are good preserved and compared to fresh tissue (a). Vessels shows a quite good preservation using PP (e) or quartz (f), even if in PP it is observed a significant lower amount of well-preserved vessels compared to fresh tissue (d). It is observed in follicles a good preservation after cryopreservation in both devices, even if quartz (i) has a significant high good preservation compared to PP (h). Follicles stages show in G two primary follicles, in H one intermediary, and in I a secondary follicle. (Magnification from 100× to 400×)
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Fig2: Light microscopy images of cortical human ovarian tissue stained with haematoxylin-eosin from fresh (control) tissue (a, d, g), cryopreserved in PP device (b, e, h) and cryopreserved in quartz device (c, f, i). Stromal cells in cryopreserved tissue using PP (b) or quartz (c) vial are good preserved and compared to fresh tissue (a). Vessels shows a quite good preservation using PP (e) or quartz (f), even if in PP it is observed a significant lower amount of well-preserved vessels compared to fresh tissue (d). It is observed in follicles a good preservation after cryopreservation in both devices, even if quartz (i) has a significant high good preservation compared to PP (h). Follicles stages show in G two primary follicles, in H one intermediary, and in I a secondary follicle. (Magnification from 100× to 400×)

Mentions: Similarly, the morphology of vessels was preserved (score 1 and 2) in all cryopreserved tissue with no statistically significant differences (PP vials 45.0 %, and quartz vials 63.6 %, resp.; p = 0.54). Results are obtained by HE staining as shown in Fig. 2.Fig. 2


Comparison of quartz vials with polypropylene vials for rapid cryopreservation of human ovarian tissue
Light microscopy images of cortical human ovarian tissue stained with haematoxylin-eosin from fresh (control) tissue (a, d, g), cryopreserved in PP device (b, e, h) and cryopreserved in quartz device (c, f, i). Stromal cells in cryopreserved tissue using PP (b) or quartz (c) vial are good preserved and compared to fresh tissue (a). Vessels shows a quite good preservation using PP (e) or quartz (f), even if in PP it is observed a significant lower amount of well-preserved vessels compared to fresh tissue (d). It is observed in follicles a good preservation after cryopreservation in both devices, even if quartz (i) has a significant high good preservation compared to PP (h). Follicles stages show in G two primary follicles, in H one intermediary, and in I a secondary follicle. (Magnification from 100× to 400×)
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC5037623&req=5

Fig2: Light microscopy images of cortical human ovarian tissue stained with haematoxylin-eosin from fresh (control) tissue (a, d, g), cryopreserved in PP device (b, e, h) and cryopreserved in quartz device (c, f, i). Stromal cells in cryopreserved tissue using PP (b) or quartz (c) vial are good preserved and compared to fresh tissue (a). Vessels shows a quite good preservation using PP (e) or quartz (f), even if in PP it is observed a significant lower amount of well-preserved vessels compared to fresh tissue (d). It is observed in follicles a good preservation after cryopreservation in both devices, even if quartz (i) has a significant high good preservation compared to PP (h). Follicles stages show in G two primary follicles, in H one intermediary, and in I a secondary follicle. (Magnification from 100× to 400×)
Mentions: Similarly, the morphology of vessels was preserved (score 1 and 2) in all cryopreserved tissue with no statistically significant differences (PP vials 45.0 %, and quartz vials 63.6 %, resp.; p = 0.54). Results are obtained by HE staining as shown in Fig. 2.Fig. 2

View Article: PubMed Central - PubMed

ABSTRACT

Background: Because higher survival of follicles during the freezing/thawing procedure improves the quality of cryopreserved tissue reimplanted after oncological therapies, defining an optimal method for human ovarian tissue cryopreservation remains a major issue in this field. One option to improve the cryopreservation procedure is to use better materials, i.e., vials with better conductivity. The aim of this study was to compare polypropylene (PP) with quartz vials. Between September 2012 and January 2013, eight patients were recruited. The ovarian cortex was cut into 3 slices, assigned randomly to a fresh and a cryopreserved group in PP (method B) or quartz vials (method C). Histological and immunohistochemical (IHC) analysis were used. For IHC three antibodies were analyzed: Ki67 (proliferation index), Bcl2 (anti apoptotic index) and Hsp70 (stress index).

Results: The majority of GCs showed positive staining for Bcl2 in both cryopreservation device, with higher expression in group C than in group B. Oocytes and their nuclei showed intense positive staining for ki67 in both methods B and C, and also a patch positive stromal cells staining for Ki67. Expression of hsp70 was not increased after cryopreservation.

Conclusions: Cryopreservation using quartz vials led to larger numbers of good follicles while maintaining consistent preservation for stromal cells and vessels.

No MeSH data available.