A microfluidic culture model of the human reprod... https://www.nature.com/articles/ncomms14584
nature nature communications articles article
Download PDF
Open Access Published: 28 March 2017
A microfluidic culture model of the human reproductive tract
and 28-day menstrual cycle
Shuo Xiao, Jonathan R. Coppeta, Hunter B. Rogers, Brett C. Isenberg, Jie Zhu, Susan A.
Olalekan, Kelly E. McKinnon, Danijela Dokic, Alexandra S. Rashedi, Daniel J. Haisenleder,
Saurabh S. Malpani, Chanel A. Arnold-Murray, Kuanwei Chen, Mingyang Jiang, Lu Bai,
Catherine T. Nguyen, Jiyang Zhang, Monica M. Laronda, Thomas J. Hope, Kruti P. Maniar,
Mary Ellen Pavone, Michael J. Avram, Elizabeth C. Sefton, Spiro Getsios, … Teresa K.
Woodruff Show authors
Nature Communications 8, Article number: 14584 (2017)
49k Accesses 249 Citations 1026 Altmetric Metrics
This article has been updated
Abstract
The endocrine system dynamically controls tissue differentiation and
homeostasis, but has not been studied using dynamic tissue culture paradigms.
Here we show that a microfluidic system supports murine ovarian follicles to
produce the human 28-day menstrual cycle hormone profile, which controls
1 of 45 10/16/22, 4:21 AM
,A microfluidic culture model of the human reprod... https://www.nature.com/articles/ncomms14584
human female reproductive tract and peripheral tissue dynamics in single, dual
and multiple unit microfluidic platforms (Solo-MFP, Duet-MFP and Quintet-
MPF, respectively). These systems simulate the in vivo female reproductive tract
and the endocrine loops between organ modules for the ovary, fallopian tube,
uterus, cervix and liver, with a sustained circulating flow between all tissues. The
reproductive tract tissues and peripheral organs integrated into a microfluidic
platform, termed EVATAR, represents a powerful new in vitro tool that allows
organ–organ integration of hormonal signalling as a phenocopy of menstrual
cycle and pregnancy-like endocrine loops and has great potential to be used in
drug discovery and toxicology studies.
Introduction
The female reproductive tract is required for the production of ova, secretion of
sex hormones and the maintenance of pregnancy throughout the gestation of
healthy offspring. Entry and advancement through puberty, normal menstrual
cycles with a potential intervening pregnancy, and endocrine support of
peripheral tissues, such as the bone, brain and heart, are all essential roles that
the reproductive tract tissues and their hormones play. The main organs of the
female reproductive tract are the ovaries, fallopian tubes, uterus and cervix.
Each organ has major responsibilities that are either autonomous (for example,
maturation of oocytes and gestation of the fetus) or interdependent such as
providing hormonal support for the tracts through which gametes (eggs and
sperm) travel and a location for the developing embryo to implant. Moreover,
each organ is composed of cells from multiple lineages (for example,
myometrium and endometrium in the uterus) that provide local function and
feedback control. Because of the intimate relationship between the cells of each
2 of 45 10/16/22, 4:21 AM
,A microfluidic culture model of the human reprod... https://www.nature.com/articles/ncomms14584
organ and between organs, toxicology studies in the female reproductive tract
have been difficult to design.
Methods used to grow mammalian cells outside the body have not
fundamentally changed in the last 50 years. Preclinical studies often begin with
individual cells, separated from cellular and physical contacts that are
important for biological function1. These dispersed cells must be propagated
through weekly reduction divisions and maintained on flat plastic; however,
these cells are missing the cell physicochemical microenvironment, three-
dimensional (3D) tissue-specific architecture, and blood flow perfusion found in
natural tissues. Furthermore, typical media composition is based on basal
nutrients, bovine serum and a few specialized factors that are placed in a static
setting with random mixing. As a consequence, cell–cell and tissue-level
cytokine and endocrine signals are not integrated into signalling pathways. In
parallel with these developments, the pharmaceutical industry is challenged by
the fact that fewer drugs are emerging to address many unmet needs, including
cardiovascular disease, cancer, immune diseases, and new contraceptives2,3,4.
Despite large investments in research funding, only ∼8% of drugs for which
Investigational New Drug applications have been filed will be approved by the
FDA5. Innovative methods to culture cells in vitro to test new compounds are
therefore necessary to reinvigorate the drug pipeline.
Recently, organ-on-a-chip and human-on-a-chip microfluidic technologies have
garnered significant interest and offer promising approaches to test the efficacy
and toxicity of new drugs in vitro16,17,18. Microfluidics represents an engineered
manipulation of fluid flow in a set of micrometre-sized channels and provides
precise control of microlitre volume of fluids. In the current study, Solo-MFP
and Duet-MFP systems based on pneumatic actuation technology and a
3 of 45 10/16/22, 4:21 AM
, A microfluidic culture model of the human reprod... https://www.nature.com/articles/ncomms14584
Quintet-MFP system based on embedded electromagnetic actuation
technology were designed for single and multiple tissue cultures, respectively.
Mouse ovarian tissue was cultured in the Solo-MFP and Duet-MFP systems for 28
days, which resulted in follicle production of 28-day menstrual cycle hormone
profiles. To test the ovarian hormone control of downstream human female
reproductive tract and peripheral tissues, the ovary, fallopian tube, uterus,
cervix and liver were cultured in the Quintet-MFP system. This integrated
microfluidic platform enables dynamic and precisely controlled interaction
between organs and is operational over the course of month-long experiments.
This represents a next step in fundamental and applied toxicology as well as
therapeutic discovery and deployment to address a wide range of biological
problems and fill the drug pipeline.
Results
Microfluidic technology enabled dynamic tissue integration
The first step of our work was to develop platforms that could sustain tissue-
level function for the length of the human menstrual cycle (that is, 28 days). The
Solo-MFP and Duet-MFP systems are based on pneumatic actuation technology,
by which the individual systems are supplied with positive and negative air
pressures via a system dock that is connected to a five-channel pressure
controller manifold (Fig. 1a,b, refer to Supplementary Table 1 for
nomenclature). The pressure of individual channels is switched between a
vacuum or pressure source using an electromagnetic three-way valve controlled
via a personalized pump programme created in LabVIEW using a computer
interface. The Solo-MFP and Duet-MFP systems use a universal pneumatic plate
that distributes positive and negative air pressures to specific valve or pump
4 of 45 10/16/22, 4:21 AM
nature nature communications articles article
Download PDF
Open Access Published: 28 March 2017
A microfluidic culture model of the human reproductive tract
and 28-day menstrual cycle
Shuo Xiao, Jonathan R. Coppeta, Hunter B. Rogers, Brett C. Isenberg, Jie Zhu, Susan A.
Olalekan, Kelly E. McKinnon, Danijela Dokic, Alexandra S. Rashedi, Daniel J. Haisenleder,
Saurabh S. Malpani, Chanel A. Arnold-Murray, Kuanwei Chen, Mingyang Jiang, Lu Bai,
Catherine T. Nguyen, Jiyang Zhang, Monica M. Laronda, Thomas J. Hope, Kruti P. Maniar,
Mary Ellen Pavone, Michael J. Avram, Elizabeth C. Sefton, Spiro Getsios, … Teresa K.
Woodruff Show authors
Nature Communications 8, Article number: 14584 (2017)
49k Accesses 249 Citations 1026 Altmetric Metrics
This article has been updated
Abstract
The endocrine system dynamically controls tissue differentiation and
homeostasis, but has not been studied using dynamic tissue culture paradigms.
Here we show that a microfluidic system supports murine ovarian follicles to
produce the human 28-day menstrual cycle hormone profile, which controls
1 of 45 10/16/22, 4:21 AM
,A microfluidic culture model of the human reprod... https://www.nature.com/articles/ncomms14584
human female reproductive tract and peripheral tissue dynamics in single, dual
and multiple unit microfluidic platforms (Solo-MFP, Duet-MFP and Quintet-
MPF, respectively). These systems simulate the in vivo female reproductive tract
and the endocrine loops between organ modules for the ovary, fallopian tube,
uterus, cervix and liver, with a sustained circulating flow between all tissues. The
reproductive tract tissues and peripheral organs integrated into a microfluidic
platform, termed EVATAR, represents a powerful new in vitro tool that allows
organ–organ integration of hormonal signalling as a phenocopy of menstrual
cycle and pregnancy-like endocrine loops and has great potential to be used in
drug discovery and toxicology studies.
Introduction
The female reproductive tract is required for the production of ova, secretion of
sex hormones and the maintenance of pregnancy throughout the gestation of
healthy offspring. Entry and advancement through puberty, normal menstrual
cycles with a potential intervening pregnancy, and endocrine support of
peripheral tissues, such as the bone, brain and heart, are all essential roles that
the reproductive tract tissues and their hormones play. The main organs of the
female reproductive tract are the ovaries, fallopian tubes, uterus and cervix.
Each organ has major responsibilities that are either autonomous (for example,
maturation of oocytes and gestation of the fetus) or interdependent such as
providing hormonal support for the tracts through which gametes (eggs and
sperm) travel and a location for the developing embryo to implant. Moreover,
each organ is composed of cells from multiple lineages (for example,
myometrium and endometrium in the uterus) that provide local function and
feedback control. Because of the intimate relationship between the cells of each
2 of 45 10/16/22, 4:21 AM
,A microfluidic culture model of the human reprod... https://www.nature.com/articles/ncomms14584
organ and between organs, toxicology studies in the female reproductive tract
have been difficult to design.
Methods used to grow mammalian cells outside the body have not
fundamentally changed in the last 50 years. Preclinical studies often begin with
individual cells, separated from cellular and physical contacts that are
important for biological function1. These dispersed cells must be propagated
through weekly reduction divisions and maintained on flat plastic; however,
these cells are missing the cell physicochemical microenvironment, three-
dimensional (3D) tissue-specific architecture, and blood flow perfusion found in
natural tissues. Furthermore, typical media composition is based on basal
nutrients, bovine serum and a few specialized factors that are placed in a static
setting with random mixing. As a consequence, cell–cell and tissue-level
cytokine and endocrine signals are not integrated into signalling pathways. In
parallel with these developments, the pharmaceutical industry is challenged by
the fact that fewer drugs are emerging to address many unmet needs, including
cardiovascular disease, cancer, immune diseases, and new contraceptives2,3,4.
Despite large investments in research funding, only ∼8% of drugs for which
Investigational New Drug applications have been filed will be approved by the
FDA5. Innovative methods to culture cells in vitro to test new compounds are
therefore necessary to reinvigorate the drug pipeline.
Recently, organ-on-a-chip and human-on-a-chip microfluidic technologies have
garnered significant interest and offer promising approaches to test the efficacy
and toxicity of new drugs in vitro16,17,18. Microfluidics represents an engineered
manipulation of fluid flow in a set of micrometre-sized channels and provides
precise control of microlitre volume of fluids. In the current study, Solo-MFP
and Duet-MFP systems based on pneumatic actuation technology and a
3 of 45 10/16/22, 4:21 AM
, A microfluidic culture model of the human reprod... https://www.nature.com/articles/ncomms14584
Quintet-MFP system based on embedded electromagnetic actuation
technology were designed for single and multiple tissue cultures, respectively.
Mouse ovarian tissue was cultured in the Solo-MFP and Duet-MFP systems for 28
days, which resulted in follicle production of 28-day menstrual cycle hormone
profiles. To test the ovarian hormone control of downstream human female
reproductive tract and peripheral tissues, the ovary, fallopian tube, uterus,
cervix and liver were cultured in the Quintet-MFP system. This integrated
microfluidic platform enables dynamic and precisely controlled interaction
between organs and is operational over the course of month-long experiments.
This represents a next step in fundamental and applied toxicology as well as
therapeutic discovery and deployment to address a wide range of biological
problems and fill the drug pipeline.
Results
Microfluidic technology enabled dynamic tissue integration
The first step of our work was to develop platforms that could sustain tissue-
level function for the length of the human menstrual cycle (that is, 28 days). The
Solo-MFP and Duet-MFP systems are based on pneumatic actuation technology,
by which the individual systems are supplied with positive and negative air
pressures via a system dock that is connected to a five-channel pressure
controller manifold (Fig. 1a,b, refer to Supplementary Table 1 for
nomenclature). The pressure of individual channels is switched between a
vacuum or pressure source using an electromagnetic three-way valve controlled
via a personalized pump programme created in LabVIEW using a computer
interface. The Solo-MFP and Duet-MFP systems use a universal pneumatic plate
that distributes positive and negative air pressures to specific valve or pump
4 of 45 10/16/22, 4:21 AM
