Autologous Co-culture of Primary Human Alveolar Macrophages and Epithelial Cells for Investigating Aerosol Medicines. Part I: Model Characterisation

Abstract

CitationAutologous co-culture of primary human alveolar macrophages and epithelial cells for investigating aerosol medicines. Part I: model characterisation. 2016, 44 (4):337-347 Altern Lab Anim Journal Alternatives to laboratory animals : ATLA Download date 10/07/2020 16:43:37 Item License http://creativecommons.org/licenses/by-nc-sa/4.0/ Link to ItemSummary -The development of new formulations for pulmonary drug delivery is a challenge on its own. New in vitro models which address the lung are aimed at predicting and optimising the quality, efficacy and safety of inhaled drugs, to facilitate the more rapid translation of such products into the clinic.Reducing the complexity of the in vivo situation requires that such models reproducibly reflect essential physiological factors in vitro. The choice of cell types, culture conditions and the experimental set-up, can affect the outcome and the relevance of a study. In the alveolar space of the lung, epithelial cells and alveolar macrophages are the most important cell types, forming an efficient cellular barrier to aerosols. Our aim was to mimic this barrier with primary human alveolar cells. Cell densities of alveolar macrophages and epithelial cells, isolated from the same human donor, were optimised, with a focus on barrier properties. The combination of 300,000 epithelial cells/cm 2 together with 100,000 macrophages/cm 2 showed a functional barrier (transepithelial electrical resistance > 500Ω.cm 2 ). This cell model was combined with the Pharmaceutical Aerosol Deposition Device on Cell Cultures. The functionality of the in vitro system was investigated with spray-dried fluorescently labelled poly(lactic-co-glycolic) acid particles loaded with ovalbumin as a model drug.

Medium throughput breathing human primary cell alveolus-on-chip model

“…7D ); the maximal TEER value of 545.5 ± 2.86 Ω cm 2 was reached at day 14. Although TEER in co-culture on our chip was smaller than previously described for hAEpCs in inserts (1000–2000 Ω cm 2 ) 80 , these values reflected a tight monolayer enabling active water and selective ion transport 73 , 77 . Differences between TEER values measured in inserts and in microfluidic systems have been reported earlier and are thought to be due to a geometrical origin rather than a biological one 81 .…”

“…The experiments could have therefore been performed for longer periods of time for defined drug discovery or toxicity assay, such as acute inhalation toxicity studies 86 . To the best of our knowledge, this is the first study in which a co-culture of primary human lung epithelial and endothelial cells was established on a lung-on-chip device in a manner that enabled elevated TEER values to be established, maintained, and measured over a long period of time 6 , 80 .…”

Short-term and Long-term Exposure of the MucilAir™ Model to Polycyclic Aromatic Hydrocarbons

“…Such models are usually a combination of target cells — for example, epithelial cells — and immune cells such as macrophages. 2,3 In view of the potential cellular interactions, coculture models can certainly provide more accurate information about the effects of chemical exposure, but the role of cell positioning and communication in the 3-D space is still not addressed in these models. In addition, the model lifespan is usually rather short, that is, days or weeks.…”

Co-culture of human alveolar epithelial (hAELVi) and macrophage (THP-1) cell lines_suppl

“…A possible alternative to mimic the human air-blood barrier are primary ATI-like cells, which express functional tight junctions when grown on permeable filter substrates (e.g., Transwells ® ) and are, therefore, better suited to evaluate transport of drugs or carriers through the epithelium (Elbert et al, 1999;Daum et al, 2012;Fuchs et al, 2003). These cells were recently combined with alveolar macrophages as an autologous co-culture (Hittinger et al, 2016). Despite the interesting aspect that both cell types originate from the same donor, such a system has several disadvantages, such as the limited supply of tissue for isolating primary cells, their short life-spans and inter-individual differences due to different donors.…”