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      A Microfluidic Device for Culturing an Encapsulated Ovarian Follicle

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          Abstract

          Microfluidic chips have been proved effective in mimicking different organs of human body. Simulating human ovarian follicles by microfluidic device will be useful in exploring the mechanism of folliculogenesis and related diseases. In this paper, a microfluidic chip was designed to culture a single human pre-antral follicle. Ovarian follicles were first encapsulated in 3D calcium alginate hydrogel beads and then cultured on chip and in dish under same conditions. The diameters of cultured ovarian follicles were measured, and the same amount of medium was collected from microfluidic device or dish per two days for measuring the estradiol and androgen concentrations. The results confirmed the successful growth of ovarian follicles on chip with their hormonal trends and diameters increase, which were similar to ovarian follicles cultured in dish. It is concluded that this microfluidic chip can be used to culture a single human ovarian follicle, which provides a useful tool to explore the hormonal changes and their interactions during folliculogenesis.

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          Most cited references32

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          A Validated Model of Serum Anti-Müllerian Hormone from Conception to Menopause

          Background Anti-Müllerian hormone (AMH) is a product of growing ovarian follicles. The concentration of AMH in blood may also reflect the non-growing follicle (NGF) population, i.e. the ovarian reserve, and be of value in predicting reproductive lifespan. A full description of AMH production up to the menopause has not been previously reported. Methodology/Principal Findings By searching the published literature for AMH concentrations in healthy pre-menopausal females, and using our own data (combined ) we have generated and robustly validated the first model of AMH concentration from conception to menopause. This model shows that 34% of the variation in AMH is due to age alone. We have shown that AMH peaks at age 24.5 years, followed by a decline to the menopause. We have also shown that there is a neonatal peak and a potential pre-pubertal peak. Our model allows us to generate normative data at all ages. Conclusions/Significance These data highlight key inflection points in ovarian follicle dynamics. This first validated model of circulating AMH in healthy females describes a transition period in early adulthood, after which AMH reflects the progressive loss of the NGF pool. The existence of a neonatal increase in gonadal activity is confirmed for females. An improved understanding of the relationship between circulating AMH and age will lead to more accurate assessment of ovarian reserve for the individual woman.
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            Tissue-engineered follicles produce live, fertile offspring.

            Oocytes grown in vitro are of low quality and yield few live births, thus limiting the ability to store or bank the ova of women wishing to preserve their fertility. We applied tissue engineering principles to the culture of immature mouse follicles by designing an alginate hydrogel matrix to maintain the oocyte's 3- dimensional (3D) architecture and cell-cell interactions in vitro. A 3D culture mimics the in vivo follicle environment, and hydrogel-encapsulated follicles develop mature oocytes within the capacity for fertilization similar to that of oocytes matured in vivo. Embryos derived from cultured oocytes fertilized in vitro and transferred to pseudopregnant female mice were viable, and both male and female offspring were fertile. Our results demonstrate that alginate hydrogel-based 3D in vitro culture of follicles permits normal growth and development of follicles and oocytes. This system creates new opportunities for discovery in follicle biology and establishes a core technology for human egg banks for preservation of fertility.
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              Aromatase expression in the ovary: hormonal and molecular regulation.

              Estrogens are synthesized by the aromatase enzyme encoded by the Cyp19a1 gene, which contains an unusually large regulatory region. In most mammals, aromatase expression is under the control of two distinct promoters a gonad- and a brain-specific promoter. In humans, this gene contains 10 tissue-specific promoters that are alternatively used in various cell types and tumors. Each promoter is regulated by a distinct set of regulatory sequences and transcription factors that bind to these specific sequences. The cAMP/PKA/CREB pathway is considered to be the primary signaling cascade through which the gonad Cyp19 promoter is regulated. Very interestingly, in rat luteal cells, the proximal promoter is not controlled in a cAMP dependent manner. Strikingly, these cells express aromatase at high levels similar to those found in preovulatory follicles, suggesting that alternative and powerful mechanisms control aromatase expression in luteal cells and that the rat corpus luteum represents an important paradigm for understanding alternative controls of the aromatase gene. Here, the molecular and cellular mechanisms controlling the expression of the aromatase gene in granulosa and luteal cells are discussed.
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                Author and article information

                Journal
                Micromachines (Basel)
                Micromachines (Basel)
                micromachines
                Micromachines
                MDPI
                2072-666X
                20 November 2017
                November 2017
                : 8
                : 11
                : 335
                Affiliations
                [1 ]Department of Biomedical Engineering, Dalian University of Technology, Dalian 116024, China; azizjatoi@ 123456hotmail.com (A.U.R.A.); 1352257919@ 123456mail.dlut.edu.cn (C.G.)
                [2 ]Dalian Institute of Maternal and Child Health Care, Dalian 116024, China; fmjnewlife@ 123456126.com
                [3 ]School of Pharmaceutical Science and Technology, Dalian University of Technology, Dalian 116024, China; dengjiu@ 123456mail.dlut.edu.cn (J.D.); yluo@ 123456dlut.edu.cn (Y.L.); bclin@ 123456dicp.ac.cn (Bi.L.)
                Author notes
                [* ]Correspondences: yuxiaohui369@ 123456163.com (X.Y.); lbo@ 123456dlut.edu.cn (Bo.L.); Tel.: +86-411-8455-2999 (X.Y.); +86-411-8470-6002 (Bo.L.)
                [†]

                These authors contributed equally to this work.

                Article
                micromachines-08-00335
                10.3390/mi8110335
                6190016
                30400524
                b9ad61dc-6ac5-4f76-a174-f79ed5b8b1a3
                © 2017 by the authors.

                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/).

                History
                : 04 October 2017
                : 17 November 2017
                Categories
                Article

                ovarian follicles,microfluidics,organ on a chip
                ovarian follicles, microfluidics, organ on a chip

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