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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...

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Autores principales: 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.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2021
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.
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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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