Quality assessment of a placental perfusion protocol

Mathiesen, Line; Mose, Tina; Mørck, Thit Juul; Nielsen, Jeanette K.; Nielsen, Leif Kofoed; Maroun, Lisa Leth; Dziegiel, Morten Hanefeld; Larsen, Lise Grupe; Knudsen, Lisbeth E.

doi:10.1016/j.reprotox.2010.01.006

Cited by 103 publications

(80 citation statements)

References 26 publications

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“…The protocol has evolved over time and can vary between different research groups especially concerning the flow rate, composition of perfusion medium, form of oxygenation and heating 26,27 . Especially the flow rate can influence the time at which transplacental transfer occurs.…”

Section: Discussionmentioning

confidence: 99%

“…To control this, the addition of a passively transported reference compound like antipyrine is important. The transfer rate of the xenobiotic can be always compared to the transfer rate of antipyrine (F/M ratio should be above 0.75) 26 . Since the antipyrine transfer is mainly limited by the flow and exchange surface, this comparison takes differences in the flow and the size of the perfused cotyledon into account which could vary between the experiments.…”

Section: Discussionmentioning

confidence: 99%

“…Since the antipyrine transfer is mainly limited by the flow and exchange surface, this comparison takes differences in the flow and the size of the perfused cotyledon into account which could vary between the experiments. In addition, FITC-dextran could be added to the fetal circuit to serve as control for the integrity of the barrier 26 . Fetal volume loss is also used as marker for the barrier integrity.…”

Section: Discussionmentioning

confidence: 99%

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Determination of the Transport Rate of Xenobiotics and Nanomaterials Across the Placenta using the <em>ex vivo</em> Human Placental Perfusion Model

Grafmüller

Manser

Krug

et al. 2013

JoVE

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Decades ago the human placenta was thought to be an impenetrable barrier between mother and unborn child. However, the discovery of thalidomide-induced birth defects and many later studies afterwards proved the opposite. Today several harmful xenobiotics like nicotine, heroin, methadone or drugs as well as environmental pollutants were described to overcome this barrier. With the growing use of nanotechnology, the placenta is likely to come into contact with novel nanoparticles either accidentally through exposure or intentionally in the case of potential nanomedical applications. Data from animal experiments cannot be extrapolated to humans because the placenta is the most species-specific mammalian organ 1 . Therefore, the ex vivo dual recirculating human placental perfusion, developed by Panigel et al. in 1967 2 and continuously modified by Schneider et al. in 1972 3 , can serve as an excellent model to study the transfer of xenobiotics or particles.Here, we focus on the ex vivo dual recirculating human placental perfusion protocol and its further development to acquire reproducible results.The placentae were obtained after informed consent of the mothers from uncomplicated term pregnancies undergoing caesarean delivery. The fetal and maternal vessels of an intact cotyledon were cannulated and perfused at least for five hours. As a model particle fluorescently labelled polystyrene particles with sizes of 80 and 500 nm in diameter were added to the maternal circuit. The 80 nm particles were able to cross the placental barrier and provide a perfect example for a substance which is transferred across the placenta to the fetus while the 500 nm particles were retained in the placental tissue or maternal circuit. The ex vivo human placental perfusion model is one of few models providing reliable information about the transport behavior of xenobiotics at an important tissue barrier which delivers predictive and clinical relevant data. Video LinkThe video component of this article can be found at

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

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Determination of the Transport Rate of Xenobiotics and Nanomaterials Across the Placenta using the <em>ex vivo</em> Human Placental Perfusion Model

Grafmüller

Manser

Krug

et al. 2013

JoVE

View full text Add to dashboard Cite

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“…6 Several experimental models have been implemented to study the disposition of substances across the human placenta, including cell lines, primary cultured cells, villous explants, isolated membrane vesicles, and the dually perfused placental cotyledon. [7][8][9] The trophoblast cell layer is the rate-limiting barrier for drug and nutrient exchange between mother and fetus, and the human choriocarcinoma trophoblastic BeWo cell line is an established in vitro model for placental transport studies of soluble material. 10 Although this model does not constitute a complete physiological system and the entire tissue microenvironment found in humans in vivo, it does represent the rate-limiting barrier of maternal-fetal exchange, irrespective of the transport mechanism through the cell layers.…”

Section: Introductionmentioning

confidence: 99%

In vitro placental model optimization for nanoparticle transport studies

Cartwright¹,

Poulsen²,

Nielsen³

et al. 2012

IJN

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Advances in biomedical nanotechnology raise hopes in patient populations but may also raise questions regarding biodistribution and biocompatibility, especially during pregnancy. Special consideration must be given to the placenta as a biological barrier because a pregnant woman's exposure to nanoparticles could have significant effects on the fetus developing in the womb. Therefore, the purpose of this study is to optimize an in vitro model for characterizing the transport of nanoparticles across human placental trophoblast cells. The growth of BeWo (clone b30) human placental choriocarcinoma cells for nanoparticle transport studies was characterized in terms of optimized Transwell® insert type and pore size, the investigation of barrier properties by transmission electron microscopy, tight junction staining, transepithelial electrical resistance, and fluorescein sodium transport. Following the determination of nontoxic concentrations of fluorescent polystyrene nanoparticles, the cellular uptake and transport of 50 nm and 100 nm diameter particles was measured using the in vitro BeWo cell model. Particle size measurements, fluorescence readings, and confocal microscopy indicated both cellular uptake of the fluorescent polystyrene nanoparticles and the transcellular transport of these particles from the apical (maternal) to the basolateral (fetal) compartment. Over the course of 24 hours, the apparent permeability across BeWo cells grown on polycarbonate membranes (3.0 µm pore size) was four times higher for the 50 nm particles compared with the 100 nm particles. The BeWo cell line has been optimized and shown to be a valid in vitro model for studying the transplacental transport of nanoparticles. Fluorescent polystyrene nanoparticle transport was size-dependent, as smaller particles reached the basal (fetal) compartment at a higher rate

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“…Of these two models, the perfused human placenta is the better representation of the in vivo situation, as it retains the complete set of placental organization. Nevertheless, it is a time-consuming method requiring fresh placental tissue, and many experiments are nullified due to high leakage and failure rates (Mathiesen et al, 2010). While perfusions with fullterm placentas cannot provide data regarding early gestation, placentas obtained following preterm deliveries may not represent normal tissue (Nanovskaya et al, 2008).…”

mentioning

confidence: 99%

Exploring the interactions of nanoparticles with multiple models of the maternal--fetal interface

Rytting

2015

Nanotoxicology

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Quality assessment of a placental perfusion protocol

Cited by 103 publications

References 26 publications

Determination of the Transport Rate of Xenobiotics and Nanomaterials Across the Placenta using the <em>ex vivo</em> Human Placental Perfusion Model

Determination of the Transport Rate of Xenobiotics and Nanomaterials Across the Placenta using the <em>ex vivo</em> Human Placental Perfusion Model

In vitro placental model optimization for nanoparticle transport studies

Exploring the interactions of nanoparticles with multiple models of the maternal--fetal interface

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