Long-term viability and differentiation of bovine oviductal monolayers: Bidimensional versus three-dimensional culture

Gualtieri, Roberto; Mollo, Valentina; Braun, Sabrina; Barbato, Vincenza; Fiorentino, Ilaria; Talevi, Riccardo

doi:10.1016/j.theriogenology.2012.06.010

Cited by 27 publications

(22 citation statements)

References 20 publications

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“…3) can be created since the medium in the culture dish is separated from the medium in the cell insert, resulting in a basolateral (petri dish) and an apical (cell insert) compartment. OECs can be cultured to confluence on the cell insert and by removing the medium in the insert an air–liquid interface is created that induces the OECs to establish polarity comparable to that seen in situ in the oviduct and to differentiate into active secretory and ciliated cells 13,14,34,39,77,90,93Figure 3Porous membrane cell culture inserts.…”

Section: Studying Oviduct Functionmentioning

confidence: 99%

“…4a). 2-D culture of OECs is hampered by a rapid loss of typical differentiated OEC properties, such as ciliation, columnar cell morphology, cell polarity, secretory granules and bulbous protrusions 8,39,40,47,48,101,104,115. The use of 2-D culture was nevertheless a useful first step in trying to understand the roles of the oviduct during gamete interaction and early embryo development.…”

Section: Approaches To Study Oviduct Functionmentioning

confidence: 99%

See 1 more Smart Citation

Designing 3-Dimensional In Vitro Oviduct Culture Systems to Study Mammalian Fertilization and Embryo Production

et al. 2016

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The oviduct was long considered a largely passive conduit for gametes and embryos. However, an increasing number of studies into oviduct physiology have demonstrated that it specifically and significantly influences gamete interaction, fertilization and early embryo development. While oviduct epithelial cell (OEC) function has been examined during maintenance in conventional tissue culture dishes, cells seeded into these two-dimensional (2-D) conditions suffer a rapid loss of differentiated OEC characteristics, such as ciliation and secretory activity. Recently, three-dimensional (3-D) cell culture systems have been developed that make use of cell inserts to create basolateral and apical medium compartments with a confluent epithelial cell layer at the interface. Using such 3-D culture systems, OECs can be triggered to redevelop typical differentiated cell properties and levels of tissue organization can be developed that are not possible in a 2-D culture. 3-D culture systems can be further refined using new micro-engineering techniques (including microfluidics and 3-D printing) which can be used to produce ‘organs-on-chips’, i.e. live 3-D cultures that bio-mimic the oviduct. In this review, concepts for designing bio-mimic 3-D oviduct cultures are presented. The increased possibilities and concomitant challenges when trying to more closely investigate oviduct physiology, gamete activation, fertilization and embryo production are discussed.

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Section: Studying Oviduct Functionmentioning

confidence: 99%

Section: Approaches To Study Oviduct Functionmentioning

confidence: 99%

Designing 3-Dimensional In Vitro Oviduct Culture Systems to Study Mammalian Fertilization and Embryo Production

et al. 2016

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“…However, up until now, the underlying causes to these observations remain unknown and studying oviduct physiology remains straining to perform [23][24][25]. In vitro research for morphologic [24,[26][27][28] and functional [29][30][31][32] characteristics of bovine oviductal epithelial cells (BOECs) has provided us with three different, well-established cell culture systems allowing the acquisition of mechanistic insights and observation of cellular pathways: (1) monolayers grown at the bottom of culture plates [33], (2) a BOEC suspension system with a high maintenance of physiological cellular characteristics [34][35][36], or (3) in hanging inserts [37][38][39] of a polarized cell culture (PCC) system allowing bidirectional cell exposure.…”

Section: Introductionmentioning

confidence: 99%

Elevated non-esterified fatty acid concentrations hamper bovine oviductal epithelial cell physiology in three different in vitro culture systems

et al. 2015

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Elevated non-esterified fatty acids (NEFAs) have been recognized as an important link between lipolytic metabolic conditions and impaired fertility in high-yielding dairy cows. However, NEFA effects on the oviductal micro-environment currently remain unknown. We hypothesize that elevated NEFAs may contribute to the complex pathology of subfertility by exerting a negative effect on bovine oviductal epithelial cell (BOEC) physiology. Therefore, the objectives of this study were to elucidate direct NEFA effects on BOEC physiology in three different in vitro cell culture systems. Bovine oviductal epithelial cells (four replicates) were mechanically isolated, pooled, and cultured as conventional monolayers, as explants, and in a polarized cell culture system with Dulbecco's modified Eagle's medium/F12-based culture medium. Bovine oviductal epithelial cells were exposed to an NEFA mixture of oleic, stearic, and palmitic acids for 24 hours at both physiological and pathologic concentrations. A control (0 μM NEFA) and a solvent control (0 μM NEFA + 0.45% ethanol) group were implemented. Bovine oviductal epithelial cells physiology was assessed by means of cell number and viability, a sperm binding assay, transepithelial electric resistance (TER), and a wound-healing assay. Bovine oviductal epithelial cell morphology was assessed by scanning electron microscopy on cell polarity, presence of microvilli and cilia, and monolayer integrity. Bovine oviductal epithelial cell number was negatively affected by increasing NEFAs, however, cell viability was not. Sperm binding affinity significantly decreased with increasing NEFAs and tended (P = 0.051) to be more affected by the direction of NEFA exposure in the polarized cell culture system. The absolute TER increase after NEFA exposure in the control (110 ± 11 Ω.cm(2)) was significantly higher than that in all the other treatments and was also different depending on the exposure side. Bidirectional exposed monolayers were even associated with a significant TER reduction (-15 ± 10 Ω.cm(2); P < 0.05). Cell proliferation capacity showed a decreased cell migration with increasing NEFA concentrations but was irrespective of the exposure side. Bovine oviductal epithelial cell morphology was not affected. In conclusion, in an in vitro setting, NEFAs exert a negative effect on BOEC physiology but not morphology. Ultimately, these physiological alterations in its microenvironment may result in suboptimal development of the pre-implantation embryo and a reduced reproductive outcome in dairy cattle.

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“…The culture system consists of an apical and a basal compartment, which are separated by a porous filter support and the confluent epithelial tissue. Upon differentiation, epithelial cells of mucociliary phenotype secrete mucus fluid into the apical compartment TA B L E 1 Examples for cell culture models of female reproductive tract epithelia using the air-liquid interface (ALI) approach Chen et al (2013aChen et al ( ,b, 2015Chen et al ( , 2018Chen et al ( , 2017 and Miessen, Sharbati, Einspanier, and Schoen (2011) Cattle Morphology, TEER, apical fluid proteome, fertilization, embryo development, embryonic marker gene expression after co-culture Chen et al (2017), Ferraz et al (2017a) and Gualtieri et al (2012Gualtieri et al ( , 2013 Mouse Morphology, TEER, apical fluid proteome, embryo development Chen et al (2017) Abbreviation: TEER, transepithelial electrical resistance.…”

Section: Morphological Appearancementioning

confidence: 99%

Air‐liquid interface cell culture: From airway epithelium to the female reproductive tract

Chen

Schoen

2019

Reprod Domestic Animals

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The air‐liquid interface (ALI) approach is primarily used to mimic respiratory tract epithelia in vitro. It is also known to support excellent differentiation of 3D multilayered skin models. To establish an ALI culture, epithelial cells are seeded into compartmentalized culture systems on porous filter supports or gel substrata. After an initial propagation period, the culture medium is removed from the apical side of the epithelium, exposing the cells to the surrounding air. Therefore, nutritive supply to the cells is warranted only by the basolateral cell pole. Under these conditions, the epithelial cells differentiate and regain full baso‐apical polarity. Some types of epithelia even generate in vivo‐like apical fluid or mucus. Interestingly, the ALI culture approach has also been shown to support morphological and functional differentiation of epithelial cells that are not normally exposed to ambient air in vivo. This review aims at giving a brief overview on the characteristics of ALI cultures in general and ALI models of female reproductive tract epithelia in particular. We discuss the applicability of ALI models for the investigation of the early embryonic microenvironment and for its implications in assisted reproductive technologies.

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Long-term viability and differentiation of bovine oviductal monolayers: Bidimensional versus three-dimensional culture

Cited by 27 publications

References 20 publications

Designing 3-Dimensional In Vitro Oviduct Culture Systems to Study Mammalian Fertilization and Embryo Production

Designing 3-Dimensional In Vitro Oviduct Culture Systems to Study Mammalian Fertilization and Embryo Production

Elevated non-esterified fatty acid concentrations hamper bovine oviductal epithelial cell physiology in three different in vitro culture systems

Air‐liquid interface cell culture: From airway epithelium to the female reproductive tract

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