2020
DOI: 10.1111/rda.13597
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Granulosa cells in three‐dimensional culture: A follicle‐like structure for domestic cat vitrified oocytes

Abstract: Despite being valuable resources for fertility preservation purposes of wild and endangered felids, cat (Felis silvestris catus) vitrified oocytes (VOs) poorly achieve full maturation after warming because their intrinsic competence is severely impaired by unavoidable cryodamages that also involve their surrounding cumulus cells (Luvoni, 2006). Modifications to media composition or supplementation with supporting cells alone have not led to truly satisfactory results, and therefore, since physical factors can … Show more

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Cited by 9 publications
(14 citation statements)
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“…This might also be related to our previous observations of VOs post-warming degeneration. Survival rates after warming of oocytes vitrified with this protocol are generally higher than 90%, but their viability decreases during the following incubation (40), as also reported in pig Cryotopvitrified oocytes (24). In other species, several studies witnessed the involvement of apoptotic cell death in VOs post-warming degeneration (10,13,14).…”
Section: Discussionsupporting
confidence: 71%
See 1 more Smart Citation
“…This might also be related to our previous observations of VOs post-warming degeneration. Survival rates after warming of oocytes vitrified with this protocol are generally higher than 90%, but their viability decreases during the following incubation (40), as also reported in pig Cryotopvitrified oocytes (24). In other species, several studies witnessed the involvement of apoptotic cell death in VOs post-warming degeneration (10,13,14).…”
Section: Discussionsupporting
confidence: 71%
“…To overcome the poor in vitro maturation and embryonic developmental rates of cat VOs, several studies focused on the modification of vitrification procedures ( 36 , 37 ), most recently with the addition of follicular fluid extracellular vesicles to vitrification-warming media ( 38 ), or on the modification of post-warming conditions, such as in vitro culture environments ( 39 , 40 ). In addition, targeting apoptosis pathways seemed to be beneficial for cat VOs developmental competence ( 4 ), and the use of apoptosis inhibitors was successful for vitrified embryos ( 20 , 21 ) and oocytes ( 14 ) in other species.…”
Section: Discussionmentioning
confidence: 99%
“…In gametes vitrified-warmed following this protocol, the most frequent morphological abnormalities are changes in ooplasm shape and granulation, partial (or, rarely, total) loss of cumulus cells and (rarely) zona pellucida fractures. On the other hand, as reported in our previous works on minimum volume vitrification with the same support, these vitrified COCs can mature 17 and develop into embryos in vitro 16 , even if at lower rates than fresh COCs. In addition, they do not usually present zona pellucida hardening (which is another well-known consequence of cryopreservation), since in vitro fertilization (IVF) is successful 16 .…”
Section: Representative Resultssupporting
confidence: 80%
“…Following cat oocyte vitrification and warming according to the present protocol (Figure 1 and Supplemental Figure 1), the vast majority of gametes survive. After vitrification, among other techniques, viability can be evaluated at the optical microscope as morphological integrity 22 1, which depicts post-warming data of oocytes intended for in vitro maturation 17 or in vitro embryo production 16 . On the whole, in the two experiments used as examples here 16 , 17 , 395 out of 435 oocytes survived, scoring an overall 90.8% postwarming viability.…”
Section: Representative Resultsmentioning
confidence: 99%
“…Only grade I oocytes [completely surrounded by more than five layers of compacted cumulus cells and with a homogeneous, dark ooplasm; ( 36 )] were selected ( n = 141). Oocytes were in vitro matured for 24 h in a controlled atmosphere (38.5°C and 5% CO 2 in air) in 100 μl microdrops of maturation medium [medium 199 supplemented with 3 mg/ml of BSA, 10 ng/ml of epidermal growth factor (EGF), 0.6 mM cysteine, and 0.5 IU/ml follicle-stimulating hormone + 0.5 IU/ml luteinizing hormone (Pluset®, Calier, Spain)] ( 37 ) covered by mineral oil in Petri dishes. At the end of IVM, oocytes were observed under the microscope to assess cumulus expansion and they were prepared for ICSI.…”
Section: Methodsmentioning
confidence: 99%