Limits...
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.


Representative images of immunohistochemical analysis of Bcl2, Ki67 and HSP70 levels in cortical human ovarian tissue. a microphotography shows Bcl2 staining in fresh stromal cells, compared to cryopreservation tissue in PP (b) and quartz (c). For follicles a positive Bcl2 staining was found in GCs after cryopreservation in PP (d) and quartz (e). Vessels evaluation shows a positive staining for Bcl2 both in fresh tissue (f) and cryopreserved tissue using PP (h) or quartz vial (g). For ki67 a positive staining after cryopreservation is shown in the nuclei like in the ooplasm using PP vial (i) and quartz vial (l). For hsp70 a diffuse positive staining in stromal cells is shows in fresh (m) and cryopreserved tissue using PP (n) or quartz devices (o). Cells stained positive for Bcl2, Ki67, HSP70 are brown, compared with the blue counterstained cells. Magnification from 25× to 400×
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

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

Fig3: Representative images of immunohistochemical analysis of Bcl2, Ki67 and HSP70 levels in cortical human ovarian tissue. a microphotography shows Bcl2 staining in fresh stromal cells, compared to cryopreservation tissue in PP (b) and quartz (c). For follicles a positive Bcl2 staining was found in GCs after cryopreservation in PP (d) and quartz (e). Vessels evaluation shows a positive staining for Bcl2 both in fresh tissue (f) and cryopreserved tissue using PP (h) or quartz vial (g). For ki67 a positive staining after cryopreservation is shown in the nuclei like in the ooplasm using PP vial (i) and quartz vial (l). For hsp70 a diffuse positive staining in stromal cells is shows in fresh (m) and cryopreserved tissue using PP (n) or quartz devices (o). Cells stained positive for Bcl2, Ki67, HSP70 are brown, compared with the blue counterstained cells. Magnification from 25× to 400×

Mentions: The majority of GCs showed positive staining for Bcl2 in both cryopreservation methods (B and C), with higher expression in group C (quartz vials) than in group B (PP vials) (Fig. 3). Oocytes and their nuclei showed intense positive staining for ki67 in both methods B and C (Fig. 3).Fig. 3


Comparison of quartz vials with polypropylene vials for rapid cryopreservation of human ovarian tissue
Representative images of immunohistochemical analysis of Bcl2, Ki67 and HSP70 levels in cortical human ovarian tissue. a microphotography shows Bcl2 staining in fresh stromal cells, compared to cryopreservation tissue in PP (b) and quartz (c). For follicles a positive Bcl2 staining was found in GCs after cryopreservation in PP (d) and quartz (e). Vessels evaluation shows a positive staining for Bcl2 both in fresh tissue (f) and cryopreserved tissue using PP (h) or quartz vial (g). For ki67 a positive staining after cryopreservation is shown in the nuclei like in the ooplasm using PP vial (i) and quartz vial (l). For hsp70 a diffuse positive staining in stromal cells is shows in fresh (m) and cryopreserved tissue using PP (n) or quartz devices (o). Cells stained positive for Bcl2, Ki67, HSP70 are brown, compared with the blue counterstained cells. Magnification from 25× to 400×
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

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

Fig3: Representative images of immunohistochemical analysis of Bcl2, Ki67 and HSP70 levels in cortical human ovarian tissue. a microphotography shows Bcl2 staining in fresh stromal cells, compared to cryopreservation tissue in PP (b) and quartz (c). For follicles a positive Bcl2 staining was found in GCs after cryopreservation in PP (d) and quartz (e). Vessels evaluation shows a positive staining for Bcl2 both in fresh tissue (f) and cryopreserved tissue using PP (h) or quartz vial (g). For ki67 a positive staining after cryopreservation is shown in the nuclei like in the ooplasm using PP vial (i) and quartz vial (l). For hsp70 a diffuse positive staining in stromal cells is shows in fresh (m) and cryopreserved tissue using PP (n) or quartz devices (o). Cells stained positive for Bcl2, Ki67, HSP70 are brown, compared with the blue counterstained cells. Magnification from 25× to 400×
Mentions: The majority of GCs showed positive staining for Bcl2 in both cryopreservation methods (B and C), with higher expression in group C (quartz vials) than in group B (PP vials) (Fig. 3). Oocytes and their nuclei showed intense positive staining for ki67 in both methods B and C (Fig. 3).Fig. 3

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.