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Capitalizing on transcriptome profiling to optimize and identify targets for promoting early murine folliculogenesis in vitro

In vitro ovarian follicle culture is an active area of research towards providing fertility options for survivors of childhood cancer. Late-stage murine follicles (multilayer secondary and onwards) can be cultured successfully to maturity to obtain a meiotically competent oocyte for fertilization, b...

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Autores principales: Jones, Andrea, Bernabé, Beatriz Peñalver, Padmanabhan, Vasantha, Li, Jun, Shikanov, Ariella
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group UK 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8206164/
https://www.ncbi.nlm.nih.gov/pubmed/34131220
http://dx.doi.org/10.1038/s41598-021-92036-y
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author Jones, Andrea
Bernabé, Beatriz Peñalver
Padmanabhan, Vasantha
Li, Jun
Shikanov, Ariella
author_facet Jones, Andrea
Bernabé, Beatriz Peñalver
Padmanabhan, Vasantha
Li, Jun
Shikanov, Ariella
author_sort Jones, Andrea
collection PubMed
description In vitro ovarian follicle culture is an active area of research towards providing fertility options for survivors of childhood cancer. Late-stage murine follicles (multilayer secondary and onwards) can be cultured successfully to maturity to obtain a meiotically competent oocyte for fertilization, but primordial and primary follicles usually die in culture because many key components of early follicle development are still unknown and difficult to mimic in vitro. To engineer a biomimetic three-dimensional culture system with high efficacy and reproducibility for the clinic, detailed mechanisms of early folliculogenesis must be uncovered. Previous studies have shown that primary murine follicles co-cultured in groups, in contrast to single follicles cultured in isolation, can reach preovulatory size and produce competent oocytes, but the factors accounting for the synergy of follicle co-culture are still unknown. To probe the underlying mechanisms of successful follicle co-culture, we conducted a time-course experiment for murine follicles encapsulated in 0.3% alginate hydrogels and compared between two conditions: groups of 5 (5X) versus groups of 10 (10X). For every 2 days during the course of 12 days, follicles were dissociated and somatic cells were isolated for microarray-based gene expression analysis (n = 380 follicles for 5X and n = 430 follicles for 10X). Gene activities in follicles co-cultured in larger groups (10X) had a distinct transcriptomic profile of key genes and pathways such as prolactin signaling and angiogenesis-related genes when compared to cells from follicles co-cultured in the smaller cohort (5X). To benchmark the results for follicles grown in culture, we compared our microarray data to data from murine follicles freshly isolated from the ovary at comparable stages of development previously published by Bernabé et al. Comparison of these datasets identified similarities and differences between folliculogenesis in the native microenvironment and the engineered in vitro system. A more detailed understanding of follicle growth in vitro will not only allow for better culture methods but also advance the field towards providing improved fertility options for survivors of childhood cancer.
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spelling pubmed-82061642021-06-16 Capitalizing on transcriptome profiling to optimize and identify targets for promoting early murine folliculogenesis in vitro Jones, Andrea Bernabé, Beatriz Peñalver Padmanabhan, Vasantha Li, Jun Shikanov, Ariella Sci Rep Article In vitro ovarian follicle culture is an active area of research towards providing fertility options for survivors of childhood cancer. Late-stage murine follicles (multilayer secondary and onwards) can be cultured successfully to maturity to obtain a meiotically competent oocyte for fertilization, but primordial and primary follicles usually die in culture because many key components of early follicle development are still unknown and difficult to mimic in vitro. To engineer a biomimetic three-dimensional culture system with high efficacy and reproducibility for the clinic, detailed mechanisms of early folliculogenesis must be uncovered. Previous studies have shown that primary murine follicles co-cultured in groups, in contrast to single follicles cultured in isolation, can reach preovulatory size and produce competent oocytes, but the factors accounting for the synergy of follicle co-culture are still unknown. To probe the underlying mechanisms of successful follicle co-culture, we conducted a time-course experiment for murine follicles encapsulated in 0.3% alginate hydrogels and compared between two conditions: groups of 5 (5X) versus groups of 10 (10X). For every 2 days during the course of 12 days, follicles were dissociated and somatic cells were isolated for microarray-based gene expression analysis (n = 380 follicles for 5X and n = 430 follicles for 10X). Gene activities in follicles co-cultured in larger groups (10X) had a distinct transcriptomic profile of key genes and pathways such as prolactin signaling and angiogenesis-related genes when compared to cells from follicles co-cultured in the smaller cohort (5X). To benchmark the results for follicles grown in culture, we compared our microarray data to data from murine follicles freshly isolated from the ovary at comparable stages of development previously published by Bernabé et al. Comparison of these datasets identified similarities and differences between folliculogenesis in the native microenvironment and the engineered in vitro system. A more detailed understanding of follicle growth in vitro will not only allow for better culture methods but also advance the field towards providing improved fertility options for survivors of childhood cancer. Nature Publishing Group UK 2021-06-15 /pmc/articles/PMC8206164/ /pubmed/34131220 http://dx.doi.org/10.1038/s41598-021-92036-y Text en © The Author(s) 2021 https://creativecommons.org/licenses/by/4.0/Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) .
spellingShingle Article
Jones, Andrea
Bernabé, Beatriz Peñalver
Padmanabhan, Vasantha
Li, Jun
Shikanov, Ariella
Capitalizing on transcriptome profiling to optimize and identify targets for promoting early murine folliculogenesis in vitro
title Capitalizing on transcriptome profiling to optimize and identify targets for promoting early murine folliculogenesis in vitro
title_full Capitalizing on transcriptome profiling to optimize and identify targets for promoting early murine folliculogenesis in vitro
title_fullStr Capitalizing on transcriptome profiling to optimize and identify targets for promoting early murine folliculogenesis in vitro
title_full_unstemmed Capitalizing on transcriptome profiling to optimize and identify targets for promoting early murine folliculogenesis in vitro
title_short Capitalizing on transcriptome profiling to optimize and identify targets for promoting early murine folliculogenesis in vitro
title_sort capitalizing on transcriptome profiling to optimize and identify targets for promoting early murine folliculogenesis in vitro
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8206164/
https://www.ncbi.nlm.nih.gov/pubmed/34131220
http://dx.doi.org/10.1038/s41598-021-92036-y
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