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Creating an Artificial 3-Dimensional Ovarian Follicle Culture System Using a Microfluidic System
We hypothesized that the creation of a 3-dimensional ovarian follicle, with embedded granulosa and theca cells, would better mimic the environment necessary to support early oocytes, both structurally and hormonally. Using a microfluidic system with controlled flow rates, 3-dimensional two-layer (co...
Autores principales: | , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2021
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7999445/ https://www.ncbi.nlm.nih.gov/pubmed/33806282 http://dx.doi.org/10.3390/mi12030261 |
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author | Healy, Mae W. Dolitsky, Shelley N. Villancio-Wolter, Maria Raghavan, Meera Tillman, Alexandra R. Morgan, Nicole Y. DeCherney, Alan H. Park, Solji Wolff, Erin F. |
author_facet | Healy, Mae W. Dolitsky, Shelley N. Villancio-Wolter, Maria Raghavan, Meera Tillman, Alexandra R. Morgan, Nicole Y. DeCherney, Alan H. Park, Solji Wolff, Erin F. |
author_sort | Healy, Mae W. |
collection | PubMed |
description | We hypothesized that the creation of a 3-dimensional ovarian follicle, with embedded granulosa and theca cells, would better mimic the environment necessary to support early oocytes, both structurally and hormonally. Using a microfluidic system with controlled flow rates, 3-dimensional two-layer (core and shell) capsules were created. The core consists of murine granulosa cells in 0.8 mg/mL collagen + 0.05% alginate, while the shell is composed of murine theca cells suspended in 2% alginate. Somatic cell viability tests and hormonal assessments (estradiol, progesterone, and androstenedione) were performed on days 1, 6, 13, 20, and 27. Confocal microscopy confirmed appropriate compartmentalization of fluorescently-labeled murine granulosa cells to the inner capsule and theca cells to the outer shell. Greater than 78% of cells present in capsules were alive up to 27 days after collection. Artificially constructed ovarian follicles exhibited intact endocrine function as evidenced by the production of estradiol, progesterone, and androstenedione. Oocytes from primary and early secondary follicles were successfully encapsulated, which maintained size and cellular compartmentalization. This novel microfluidic system successfully encapsulated oocytes from primary and secondary follicles, recapitulating the two-compartment system necessary for the development of the mammalian oocyte. Importantly, this microfluidic system can be easily adapted for sterile, high throughput applications. |
format | Online Article Text |
id | pubmed-7999445 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
spelling | pubmed-79994452021-03-28 Creating an Artificial 3-Dimensional Ovarian Follicle Culture System Using a Microfluidic System Healy, Mae W. Dolitsky, Shelley N. Villancio-Wolter, Maria Raghavan, Meera Tillman, Alexandra R. Morgan, Nicole Y. DeCherney, Alan H. Park, Solji Wolff, Erin F. Micromachines (Basel) Article We hypothesized that the creation of a 3-dimensional ovarian follicle, with embedded granulosa and theca cells, would better mimic the environment necessary to support early oocytes, both structurally and hormonally. Using a microfluidic system with controlled flow rates, 3-dimensional two-layer (core and shell) capsules were created. The core consists of murine granulosa cells in 0.8 mg/mL collagen + 0.05% alginate, while the shell is composed of murine theca cells suspended in 2% alginate. Somatic cell viability tests and hormonal assessments (estradiol, progesterone, and androstenedione) were performed on days 1, 6, 13, 20, and 27. Confocal microscopy confirmed appropriate compartmentalization of fluorescently-labeled murine granulosa cells to the inner capsule and theca cells to the outer shell. Greater than 78% of cells present in capsules were alive up to 27 days after collection. Artificially constructed ovarian follicles exhibited intact endocrine function as evidenced by the production of estradiol, progesterone, and androstenedione. Oocytes from primary and early secondary follicles were successfully encapsulated, which maintained size and cellular compartmentalization. This novel microfluidic system successfully encapsulated oocytes from primary and secondary follicles, recapitulating the two-compartment system necessary for the development of the mammalian oocyte. Importantly, this microfluidic system can be easily adapted for sterile, high throughput applications. MDPI 2021-03-04 /pmc/articles/PMC7999445/ /pubmed/33806282 http://dx.doi.org/10.3390/mi12030261 Text en © 2021 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) ). |
spellingShingle | Article Healy, Mae W. Dolitsky, Shelley N. Villancio-Wolter, Maria Raghavan, Meera Tillman, Alexandra R. Morgan, Nicole Y. DeCherney, Alan H. Park, Solji Wolff, Erin F. Creating an Artificial 3-Dimensional Ovarian Follicle Culture System Using a Microfluidic System |
title | Creating an Artificial 3-Dimensional Ovarian Follicle Culture System Using a Microfluidic System |
title_full | Creating an Artificial 3-Dimensional Ovarian Follicle Culture System Using a Microfluidic System |
title_fullStr | Creating an Artificial 3-Dimensional Ovarian Follicle Culture System Using a Microfluidic System |
title_full_unstemmed | Creating an Artificial 3-Dimensional Ovarian Follicle Culture System Using a Microfluidic System |
title_short | Creating an Artificial 3-Dimensional Ovarian Follicle Culture System Using a Microfluidic System |
title_sort | creating an artificial 3-dimensional ovarian follicle culture system using a microfluidic system |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7999445/ https://www.ncbi.nlm.nih.gov/pubmed/33806282 http://dx.doi.org/10.3390/mi12030261 |
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