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Native matrix-based human lung alveolar tissue model in vitro: studies of the reparatory actions of mesenchymal stem cells

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Abstract

Studies of lung diseases in vitro often rely on flat, plastic-based monocultures, due to short lifespan of primary cells, complicated anatomy, lack of explants, etc. We hereby present a native 3D model with cues for repopulating epithelial cells. Abilities of mesenchymal stem cells (MSC) to modulate bacterial lipopolysaccharide (LPS) and cigarette smoke-induced injury to pulmonary epithelium were tested in our model. Post-mortem human lung tissue was sliced, cut and decellularized. Resulting matrix pads were reseeded with pulmonary epithelium (A549 line). Markers of the layer integrity and certain secreted proteins in the presence of cigarette smoke extract (CSE) and LPS were assessed via Western blot, ELISA and RT-PCR assays. In parallel, the effects of MSC paracrine factors on exposed epithelial cells were also investigated at gene and protein levels. When cultured on native 3D matrix, A549 cells obtain dual, type I- and II-like morphology. Exposure to CSE and LPS leads to downregulation of several epithelial proteins and suppressed proliferation rate. MSC medium added to the model restores proliferation rate and some of the epithelial proteins, i.e. e-cadherin and beta-catenin. CSE also increases secretion of pro-inflammatory cytokines by epithelial cells and upregulates transcription factor NFκB. Some of these effects might be counteracted by MSC in our model. We introduce repopulated decellularized lung matrix that highly resembles in vivo situation and is convenient for studies of disease pathogenesis, cytotoxicology and for exploring therapeutic strategies in the human lung context in vitro. MSC paracrine products have produced protecting effects in our model.

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Acknowledgments

We would like to thank Dr. A. Rimkevicius for the assistance with post-mortem lung tissue collection, colleagues at National Center of Pathology, at Vilnius University Hospital Santariskiu Clinic for the excellent immunohistochemistry work and surgeons at the Dept. of Traumatology at the Republic Hospital of Vilnius University for their help with bone marrow collection.

Funding

This work was supported by European Social Fund (ESF) under the Development Action Program of Human Resources by implementation of the project “Execution of functions assigned to Lithuanian Research Council for implementation of the Global Grant measure” (Grant # VP1-3.1-SMM-07-K-03-052).

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    1. State Research Institute Centre for Innovative Medicine, Santariskiu 5, LT-08046, Vilnius, Lithuania

      Ieva Bruzauskaite, Jovile Raudoniute, Jaroslav Denkovskij, Edvardas Bagdonas, Sandra Meidute-Abaraviciene, Daiva Bironaite, Almantas Siaurys, Eiva Bernotiene & Ruta Aldonyte

    2. Department of Pathology, Vilnius City Hospital, Antakalnio 57, Vilnius, Lithuania

      Vaida Simonyte

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    1. Ieva Bruzauskaite
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    Correspondence to Ruta Aldonyte.

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    Ieva Bruzauskaite and Jovile Raudoniute have contributed equally to this work.

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    Bruzauskaite, I., Raudoniute, J., Denkovskij, J. et al. Native matrix-based human lung alveolar tissue model in vitro: studies of the reparatory actions of mesenchymal stem cells. Cytotechnology 69, 1–17 (2017). https://doi.org/10.1007/s10616-016-0021-z

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