Morphologic characterization of isolated bovine early preantral follicles during short-term individual in vitro culture

Jorssen, E.P.A.; Langbeen, A.; Marei, Waleed F.A.; Fransén, Erik; Porte, H.F.M. De; Leroy, J.L.M.R.; Bols, P.E.J.

doi:10.1016/j.theriogenology.2015.03.020

Cited by 12 publications

(8 citation statements)

References 46 publications

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“…During the 4-day culture period in this experiment, or as a previously investigated 10-day [37] culture period, our results showed no follicle attachment to the culture dish, no spreading of granulosa cells or release of the oocyte after in vitro culture of isolated bovine PAFs. However, since we only applied short-term culture periods, these features are likely to occur during longer culture periods.…”

Section: Discussionsupporting

confidence: 57%

“…As described earlier [37], adult bovine ovaries were collected upon slaughter and transported in warm physiologic saline (0.9% NaCl, Braun) to the laboratory within 3 h at 25°C. Following removal of the adnexa, the ovaries were washed in warm (38.5°C) physiologic solution supplemented with kanamycin (0.25%) and rinsed in alcohol (70%).…”

Section: Collection Of Ovaries Ovarian Follicle Isolation and Culturementioning

confidence: 99%

“…2). To evaluate follicles, the connection between the oocyte and the surrounding granulosa cells and the integrity of the basal membrane were microscopically evaluated as reported by [37] ( Fig. 1).…”

Section: Collection Of Ovaries Ovarian Follicle Isolation and Culturementioning

confidence: 99%

“…As stated previously [37] fixed follicles were placed on a glass slide. Thirty microliters of 80°C pre-heated Histogel® (Thermofisher, Breda, The Netherlands) was added to the follicles.…”

Section: Follicle Mounting and Histological Assessmentmentioning

confidence: 99%

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Effects of vitrification on the viability of alginate encapsulated isolated bovine pre-antral follicles

Bus

Hoeck

Langbeen

et al. 2018

J Assist Reprod Genet

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Our results show that, using a vitification protocol optimized for non-embedded follicles, 2D culture is more effective in vitrifying isolated follicles. However, embedding in alginate allow to handle follicles more efficiently, i.e., without excessive manipulation and thus less labor-intensive in combination with a reduced loss of follicles during the procedure. Based on the increased work efficiency, but lower viability and higher proportion of follicles showing impaired morphology, we consider it advantageous to optimize the protocol for the vitrification of embedded follicles to increase survival and maintain morphology after vitrification.

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Section: Discussionsupporting

confidence: 57%

Section: Collection Of Ovaries Ovarian Follicle Isolation and Culturementioning

confidence: 99%

Section: Collection Of Ovaries Ovarian Follicle Isolation and Culturementioning

confidence: 99%

Section: Follicle Mounting and Histological Assessmentmentioning

confidence: 99%

See 2 more Smart Citations

Effects of vitrification on the viability of alginate encapsulated isolated bovine pre-antral follicles

Bus

Hoeck

Langbeen

et al. 2018

J Assist Reprod Genet

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“…[3][4][5] In vitro follicular maturation techniques have been developed for humans, but their improvement becomes difficult due to the enormous difficulty in obtaining tissue and due to ethical issues. 12 The results of human studies are still limited to the study by Amorim et al, 13 in which pre-antral ovarian follicles survived for a period of 7 days after cryopreservation. Rodrigues et al 14 reported the first in vitro culture of fresh human pre-antral follicles using the three-dimensional system (3D) in Brazil.…”

Section: Introductionmentioning

confidence: 99%

Follicle Viability after Vitrification of Bovine Ovarian Tissue

Guedes

Rodrigues

Campos

et al. 2017

Rev Bras Ginecol Obstet

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Keywords► in vitro follicular maturation ► fertility preservation ► vitrification AbstractPurpose The present study aimed to evaluate the impact of vitrification on the viability of follicles using a three-dimensional (3D) in vitro culture. Methods Bovine ovarian tissue samples (n ¼ 5) obtained from slaughterhouses were utilized. The cortex was cut into small fragments of 2 Â 3 Â 0.5 mm using a tissue slicer. From these fragments, secondary follicles were first isolated by mechanical and enzymatic methods, then encapsulated in alginate gel and individually cultured for 20 days. Additional fragments of the same ovarian tissue were vitrified in a solution containing 25% glycerol and 25% ethylene glycol. After warming, the follicles underwent the same follicular isolation process that was performed for the fresh follicles. Results A total of 61 follicles were isolated, 51 from fresh ovarian tissue, and 10 from vitrified tissue. After the culture, the vitrified and fresh follicles showed 20% and 43.1% survival rates respectively (p ¼ 0.290), with no significant differences. At the end of the culture, there were no significant differences in follicular diameter between the vitrified (422.93 AE 85.05 µm) and fresh (412.99 AE 102.55 µm) groups (p ¼ 0.725). Fresh follicles showed higher mean rate of antrum formation when compared with vitrified follicles (47.1% and 20.0% respectively), but without significant difference (p ¼ 0.167). Conclusions The follicles were able to develop, grow and form antrum in the 3D system after vitrification, despite the lower results obtained with the fresh tissue. ResumoObjetivo O presente estudo teve como objetivo avaliar o impacto da vitrificação na viabilidade dos folículos utilizando a cultura in vitro tridimensional (3D). Métodos Foi utilizado tecido ovariano bovino (n ¼ 5) obtido de abatedouros. O córtex foi cortado em pequenos fragmentos de 2 Â 3 Â 0,5 mm, utilizando o tissue slicer e a partir destes fragmentos foram isolados folículos secundários por meio de

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Preservation of connexin 43 and transzonal projections in isolated bovine pre-antral follicles before and following vitrification

Bus

Szymanska

Pintelon

et al. 2020

J Assist Reprod Genet

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Morphologic characterization of isolated bovine early preantral follicles during short-term individual in vitro culture

Cited by 12 publications

References 46 publications

Effects of vitrification on the viability of alginate encapsulated isolated bovine pre-antral follicles

Effects of vitrification on the viability of alginate encapsulated isolated bovine pre-antral follicles

Follicle Viability after Vitrification of Bovine Ovarian Tissue

Preservation of connexin 43 and transzonal projections in isolated bovine pre-antral follicles before and following vitrification

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