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Experimental design. a Eight pieces were taken from each pair of ovaries, from which one was immediately fixed (fresh D0) and three were immediately grafted. Four fragments were cryopreserved and, after thawing, one fragment was fixed (cryo D0) and three were grafted. b Each cat had six ovarian tissue fragments grafted to the subcutaneous tissue of the dorsal neck—three fresh grafts and three cryopreserved grafts. The grafts were recovered on days 2 (D2), 4 (D4) and 6 (D6) after transplantation, one fresh and one cryopreserved at each time
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The present study evaluated revascularization time of fresh and cryopreserved cat ovarian tissue after transplantation to subcutaneous tissue. Ovaries of five cats were used and eight pieces of ovarian tissue were taken from each pair of ovaries. Immediately after removal, three pieces were transplanted and one fixed for fresh control. The remainin...
Citations
... Thus, some canid species have been classified as near threatened (Maned wolf -Chrysocyon brachyurus) or threatened (Ethiopian wolf-Canis simensis), or critically endangered (Red wolf -Canis rufus), according to the red list (IUCN 2023). Therefore, different reproductive strategies are being developed to preserve the fertility of both wild species that are at risk of extinction, as well as domestic animals of high genetic value, commercial or sentimental, using animal models (Abe et al. 2011;Johnson et al. 2014;Paz 2015;Lee et al. 2018), that can even configure applications for human reproduction (Scalercio et al. 2015;Donfack et al. 2018;Costa et al. 2020;Souza et al. 2020). ...
... Transplantation of ovarian tissue, associated with cryopreservation, has become an increasingly important tool in studies focused on folliculogenesis and reproductive biology ( (Terazono et al. 2012a;Campos-Junior et al. 2016), enabling fertility restoration (Bordes et al. 2005;Donnez et al. 2014;Tammiste et al. 2019;Hoekman et al. 2020). Among the types of transplants, autotransplantation has been showing highlights for not requiring immunosuppressive therapy, immune regulators, or knockout mice and for presenting less possibility of rejection (Costa et al. 2020). In addition, allotransplantation or xenotransplantation requires expensive techniques and specialized and equipped laboratories. ...
... The subcutaneous site has proven to be a suitable environment because it is easy to monitor and access, is low invasive , and does not require general anesthesia (Donfack et al. 2017), in addition to facilitating graft recovery. These sites are considered preferred sites for collecting oocytes to be used in in vitro fertilization (Costa et al. 2020). In subcutaneous sites, the neck region has been used to obtain favorable follicular survival rates and functional resumption of ovarian tissue after autotransplantation (Leonel et al. 2018;Vilela et al. 2019;Costa et al. 2020;Souza et al. 2020). ...
Ovarian tissue transplantation makes it possible to restore fertility; however, the success of this technique depends on the transplant region used. Therefore, this study aimed to evaluate the effect of two subcutaneous regions on canine ovarian transplantation, pinna (Pi) and neck (Ne), for 7 and 15 days. Ovaries collected by ovariosalpingohysterectomy were fragmented using a punch device. Fresh fragments were fixed, and the others were immediately grafted onto the animal itself in the Pi and Ne regions for 7 and 15 days. Recovered fragments were evaluated for histology (morphology, development and stromal density), picrosirius (collagen fibers), and immunohistochemistry (fibrosis and cell proliferation). The results showed that follicular normality rates were lower in Pi-7 (78%) vs. control (90%) and Pi-15 (86%), similar in Ne-7 (92%) and superior in Ne-15 (97%) compared to the control, with the effect of the region Ne (94%) superior (P < 0.05) to Pi (82%). Stromal density reduced in both regions vs. control but was similar within 15 days. Fragments from both regions showed higher fibronectin labeling and deposition of type I and lower type III collagen fibers (P < 0.05) vs. control. Proliferation rates in Ne-7 were higher (P < 0.05) than in control, and Pi-15 was higher (P < 0.05) than Ne-15. In conclusion, the pinna may be a region with greater potential than the neck after a 15-day autotransplantation of canine ovarian tissue.
... Thus, some canid species have been classi ed as vulnerable or "near threatened" (IUCN 2021). Therefore, different reproductive strategies are being developed to preserve the fertility of both wild species that are at risk of extinction, as well as domestic animals of high genetic value, commercial or sentimental, using animal models ( (Costa et al. 2020). In addition, allotransplantation or xenotransplantation requires expensive techniques and specialized and equipped laboratories. ...
... The subcutaneous site has proven to be a suitable environment because it is easy to monitor and access, is low invasive , and does not require general anesthesia (Donfack et al. 2017), in addition to facilitating graft recovery. These sites are considered preferred sites for collecting oocytes to be used in in vitro fertilization (Costa et al. 2020). In subcutaneous sites, the neck region has been used to obtain favorable follicular survival rates and functional resumption of ovarian tissue after autotransplantation (Leonel et Souza et al. 2020). ...
... Ovarian tissue transplantation may be a promising technique for resuming the reproductive potential of domestic species with high genetic value or endangered wild species(Terazono et al. 2012a;Leonel et al. 2018). Still, among the types of transplants, autotransplantation has stood out for preventing graft rejection and not requiring immunosuppressive therapy(Costa et al. 2020). However, the graft region has been considered essential in maintaining and functional resumption of ovarian tissue in transplantation(Cao and Li, 2019). ...
Ovarian tissue transplantation makes it possible to restore fertility; however, the success of this technique depends on the transplant region used. Therefore, this study aimed to evaluate the effect of two subcutaneous regions on canine ovarian transplantation, pinna (Pi) and neck (Ne), for 7 and 15 days. Ovaries collected by ovariosalpingohysterectomy were fragmented using a punch device. Fresh fragments were fixed, and the others were immediately grafted onto the animal itself in the Pi and Ne regions for 7 and 15 days. Recovered fragments were evaluated for histology (morphology, development and stromal density), picrosirius (collagen fibers), and immunohistochemistry (fibrosis and cell proliferation). The results showed that follicular normality rates were lower in Pi-7 (78%) vs. control (90%) and Pi-15 (86%), similar in Ne-7 (92%) and superior in Ne-15 (97%) compared to the control, with the effect of the region Ne (94%) superior (P < 0.05) to Pi (82%). Stromal density reduced in both regions vs . control but was similar within 15 days. Fragments from both regions showed higher fibronectin labeling and deposition of type I and lower type III collagen fibers (P < 0.05) vs . control. Proliferation rates in Ne-7 were higher (P < 0.05) than in control, and Pi-15 was higher (P < 0.05) than Ne-15. In conclusion, the pinna can be a potential region for canine ovarian tissue autotransplantation than the neck region in 15 days.
... Ischemia and degeneration of follicles are important factors, which should be considered during the transplantation of ovarian tissue and it can cause a decline in ovarian reserve and fertility potential (5)(6)(7)(8). The vascular condition of the graft site plays a critical role in the reduction of ischemia. ...
... The vascular condition of the graft site plays a critical role in the reduction of ischemia. Therefore, angiogenesis in transplanted tissue and growth of the ovarian follicles depends on the graft site (5,6,8). Vascular endothelial growth factor (VEGF) plays an important role in the posttransplantation angiogenesis of ovarian tissue (9)(10)(11). ...
Background: Several conflicting results have been reported on the survival and function of transplanted ovaries.
Objective: Evaluation of the follicular development and the expression of vascular endothelial growth factor (VEGF) and Bcl-2-associated X protein (BAX) in ovaries transplanted into uni- and bilaterally ovariectomized mice.
Materials and Methods: In this experimental study, 40 female NMRI mice (21-days-old, 12-15 gr) were ovariectomized uni- and bilaterally (n = 20/ group), while the 8-wk-old mice were considered as intact control group (n = 6). 5 weeks after transplantation at the proestrus stage, the morphology of recovered transplanted ovaries and the proportion of follicles were studied at different developmental stages. The apoptosis cell death by pro-apoptotic protein BAX and the expression of VEGF were evaluated using immunohistochemistry.
Results: In the bilaterally ovariectomized mice, among the 455 counted normal follicles, a lower rate of primordial and primary follicles and a higher rate of preantral and antral follicles were observed (p = 0.002). However, the percentages of preantral and antral follicles, and the corpus luteum were significantly lower in the intact control group (among the 508 counted normal follicles in this group) compared to other transplanted groups (p = 0.002). The number of BAX-positive cells in all groups was not significantly different. The VEGF expression was prominent in vessels of the corpus luteum, and also in the theca layer of large follicles of studied groups.
Conclusion: Early discharge of ovarian reserve was prominent in the bilaterally ovariectomized group but the incidence of apoptotic cells and VEGF expression as angiogenic factor did not differ in both ovariectomized mice. Thus, unilaterally ovariectomy has less side effects on the ovarian reserve compared to bilateral ovariectomy.
The aim of this study was to reveal (1) the determination of the serum antimüllerian hormone concentrations in cats with ovarian remnant syndrome and ovarian cyst, (2) the effectiveness of the serum antimüllerian hormone concentrations in detecting gonadectomy status, and (3) ovarian expression of the antimüllerian hormone in the ovarian cysts and ovarian remnant tissue. For this purpose, a total of 31 domestic cats older than 1 year, including ORS (n = 9), OC (n = 3), spayed (n = 8), and intact (n = 11), were used in the study. Serum and intrafollicular AMH concentrations were determined by the electrochemiluminescence immunoassay, and expression of the AMH was determined by the immunohistochemical method. The serum AMH concentration in the ORS group was 1.49 ± 1.24 ng/ml (0.10-3.63 ng/ml) whereas it was 1.78 ± 0.92 ng/ml (0.8-2.63 ng/ml) in the OC group. AMH concentration in the intrafollicular fluid was determined as 0.023 ng/ml in a cat with an ovarian cyst. While the serum AMH concentration was below 0.01 ng/ml in spayed cats, it was 5.80 ± 3.30 ng/ml (1.88-11.86 ng/ml) in the intact cats. There was a significant difference between the spayed and intact groups in terms of serum AMH concentration (p < 0.05). Moreover, in cats with ovarian cyst and ovarian remnant syndrome, no difference was found between the groups in terms of serum AMH concentration (p > 0.05). No difference was found between the ovarian remnant syndrome and intact cats in terms of serum AMH concentration (p > 0.05). In immunohistochemical staining, ovarian cysts were found to be negative for AMH while a moderate (+2) AMH positivity was observed in other follicles. Furthermore, in cats with ovarian remnant syndrome, severe (+3) AMH positivity was observed. As a result, it was determined that gonadectomy status can be accurately diagnosed by the AMH measurement. Ovarian remnant syndrome cannot be diagnosed accurately by AMH measurement. In addition, it was found that AMH did not seem to play a role in the pathogenesis of ovarian cysts in cats.
