ArticleJournal of assisted reproduction and genetics2014
Effects of neutral red assisted viability assessment on the cryotolerance of isolated bovine preantral follicles.
Article in Journal of assisted reproduction and genetics, 2014. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 6 papers.
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Who cites it
6 citing papers in PubMed.
- Effect of Elamipretide on the Vitrification of Mouse Ovarian Tissue by Freezing.Biopreservation and biobanking · 2024Article
- Method of Isolation and In Vitro Culture of Primordial Follicles in Bovine Animal Model.Methods in molecular biology (Clifton, N.J.) · 2024Article
- Purging human ovarian cortex of contaminating leukaemic cells by targeting the mitotic catastrophe signalling pathway.Journal of assisted reproduction and genetics · 2021Article
- Preservation of connexin 43 and transzonal projections in isolated bovine pre-antral follicles before and following vitrification.Journal of assisted reproduction and genetics · 2021Article
- Effects of vitrification on the viability of alginate encapsulated isolated bovine pre-antral follicles.Journal of assisted reproduction and genetics · 2018Article
- Development of macaque secondary follicles exposed to neutral red prior to 3-dimensional culture.Journal of assisted reproduction and genetics · 2018Article
Corrections and comments
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Authors and funding
6 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
purposeFertility preservation strategies warrant non-invasive viability assessment of preantral follicles (PAF) such as staining with Neutral Red (NR) that is incorporated by viable follicles. To optimize the procedure, we firstly determined the lowest concentration and shortest exposure time needed for optimal viability screening of isolated bovine PAF. Secondly, we combined this protocol to a vitrification procedure to assess cryotolerance of the stained follicles.
methodsIsolated PAF (900, divided over 6 replicates) were cultured in DMEM/Ham's F12 (Culture Medium - Cm) for 4 days (38.5 °C, 5% CO2). On D0, D2 and D4, follicles were stained, by adding NR medium (NRm = Cm with different concentrations NR) after which viability was assessed by counting stained/non-stained PAF every 30 min for a period of 2 h.
resultsFollowing a binary logistic regression analysis with staining as a result (yes/no) versus log-concentration, a probability model could be fitted, indicating that the proportion of stained follicles remained stable after 30 min when 15 μg/ml NR was used, without compromising follicular health and viability. Consequently, using this protocol, no significant effect of staining prior to vitrification, was found on PAF viability immediately after warming or following 4 days of culture.
conclusionsIn conclusion, we propose NR staining as a non-invasive, non-detrimental viability assessment tool for PAF, when applied at 15 μg/ml for 30 min, being perfectly compatible with PAF vitrification.
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