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CC BY-NC-ND 4.0 · Rev Bras Ginecol Obstet 2017; 39(11): 614-621
DOI: 10.1055/s-0037-1606129
Original Article
Thieme Revinter Publicações Ltda Rio de Janeiro, Brazil

Follicle Viability after Vitrification of Bovine Ovarian Tissue

Viabilidade folicular após a vitrificação de tecido ovariano bovino
Janaína de Souza Guedes
1   Clínica Pró-Criar - Reproductive Medicine, Belo Horizonte, MG, Brasil
2   Post-graduation, Faculdade de Ciências Médicas de Minas Gerais, Belo Horizonte, MG, Brasil
,
Jhenifer Kliemchen Rodrigues
3   Rede Brasileira de Oncofertilidade – Brazilian Oncofertility Consortium, Belo Horizonte, MG, Brazil
4   In Vitro Consultoria, Belo Horizonte, MG, Brazil
,
Ana Luisa Menezes Campos
1   Clínica Pró-Criar - Reproductive Medicine, Belo Horizonte, MG, Brasil
2   Post-graduation, Faculdade de Ciências Médicas de Minas Gerais, Belo Horizonte, MG, Brasil
,
Camila Cruz de Moraes
1   Clínica Pró-Criar - Reproductive Medicine, Belo Horizonte, MG, Brasil
2   Post-graduation, Faculdade de Ciências Médicas de Minas Gerais, Belo Horizonte, MG, Brasil
,
João Pedro Junqueira Caetano
1   Clínica Pró-Criar - Reproductive Medicine, Belo Horizonte, MG, Brasil
2   Post-graduation, Faculdade de Ciências Médicas de Minas Gerais, Belo Horizonte, MG, Brasil
3   Rede Brasileira de Oncofertilidade – Brazilian Oncofertility Consortium, Belo Horizonte, MG, Brazil
,
Ricardo Mello Marinho
1   Clínica Pró-Criar - Reproductive Medicine, Belo Horizonte, MG, Brasil
2   Post-graduation, Faculdade de Ciências Médicas de Minas Gerais, Belo Horizonte, MG, Brasil
3   Rede Brasileira de Oncofertilidade – Brazilian Oncofertility Consortium, Belo Horizonte, MG, Brazil
› Author Affiliations
Further Information

Publication History

22 November 2016

09 June 2017

Publication Date:
31 August 2017 (online)

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Abstract

Purpose 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 = 5) obtained from slaughterhouses were utilized. The cortex was cut into small fragments of 2 × 3 × 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 = 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 ± 85.05 µm) and fresh (412.99 ± 102.55 µm) groups (p = 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 = 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.

Resumo

Objetivo 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 = 5) obtido de abatedouros. O córtex foi cortado em pequenos fragmentos de 2 × 3 × 0,5 mm, utilizando o tissue slicer e a partir destes fragmentos foram isolados folículos secundários por meio de método enzimático e mecânico, encapsulados em gel de alginato e cultivados individualmente durante 20 dias. Outros fragmentos do mesmo tecido ovariano foram vitrificados em solução contendo 25% de glicerol e 25% de etilenoglicol. Após aquecimento, os folículos passaram pelo mesmo processo de isolamento folicular realizado a fresco.

Resultados Foram isolados 61 folículos, sendo 51 originários de tecido ovariano a fresco, e 10 de tecido vitrificado. Após a cultura, os folículos vitrificados apresentaram taxa de sobrevida de 20%, e o grupo a fresco apresentou taxa de 43,1% (p = 0,290). O diâmetro folicular ao final da cultura também não apresentou diferença significativa entre o grupo vitrificado (422,93 ± 85,05 µm) e a fresco (412,99 ± 102,55 µm) (p = 0,725). Os folículos a fresco apresentaram maior taxa média de formação de antro do que os folículos vitrificados (47,1% e 20,0%, respectivamente), mas sem diferença significativa (p = 0,167).

Conclusões Os folículos foram capazes de se desenvolver, crescer e formar antro em sistema 3D após a vitrificação.

Keywords

in vitro follicular maturation - fertility preservation - vitrification

Palavras-chave

maturação folicular in vitro - preservação da fertilidade - vitrificação
 

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