2D in vitro cell culture
- Airway smooth muscle cells, lung fibroblasts, airway epithelial cells, endothelial cells, mast cells
- All from human origin
- Analysis of anti-inflammatory efficacy and anti-remodelling efficacy
We use, amongst others, human airway smooth muscle cells, lung fibroblasts and airway epithelial cells, and analyse the anti-inflammatory and anti-remodelling efficacy of your compound. The exact assays and read-outs of choice will be discussed dependent on your needs, and can include;
Inhibition of cytokine (IL-1β, IL-13, TNF-α, others) or cigarette smoke induced responses, including:
- ELISA or Luminex analysis of pro-inflammatory cytokine release.
- Mucus seceretion (airway epithelial cells).
- Gene and protein expression for inflammatory markers such as COX-2.
- Activation of transcriptional mechanisms (e.g. STAT).
Inhibition of growth factor (TGF-β, PDGF, WNT, others) induced responses, including:
- Cell proliferation.
- Cell migration or adhesion (xCELLigence).
- Myofibroblast and smooth muscle differentiation (lung fibroblasts, airway smooth muscle).
- Epithelial mesencymal transition (EMT; epithelial cells).
- Gene and protein expression for matrix proteins, proliferation markers.
- Activation of transcriptional mechanisms (e.g. Smad, β-catenin, Sp1, others)
In addition, we can offer several possibilities for target validation in human samples. Please contact us to see how we can help you.
3D in vitro cell culture
- Air-liquid interface culture of airway epithelial cells
- Co-culture with fibroblasts or other cell types on request
Adequate study of epithelial cell function requires studies of human airway epithelial cells in air-liquid interface. Using this system, human airway epithelial cells are cultured on transwell inserts and air-exposed to allow differentiation into pseudostratified epithelia. The system is particularly suitable for the study of investigational compounds into epithelial cell differentiation, goblet cell metaplasia and epithelial barrier function. Co-cultures with fibroblasts (healthy and diseased) is possible. Typical read-outs include (immune)histochemistry, gene expression and ELISA of secreted factors in the medium. We have shown efficacy of tiotropium against IL-13 induced goblet cell metaplasia
– Kistemaker LEM, Hiemstra PS, Bos IST, Bouwman S, van den Berge M, Hylkema MN, Meurs H, Kerstjens HAM, Gosens R. Tiotropium attenuates IL-13-induced goblet cell metaplasia of human airway epithelial cells. Thorax 2015; 70:668-76.
– Oenema TA, Mensink G, Smedinga L, Halayko AJ, Zaagsma J, Meurs H, Gosens R, Dekkers BG. Cross-Talk between Transforming Growth Factor-beta1 and Muscarinic M2 Receptors Augments Airway Smooth Muscle Proliferation. Am J Respir Cell Mol Biol 2013;49:18-27.
– Baarsma HA, Spanjer AI, Haitsma G, Engelbertink LH, Meurs H, Jonker MR, Timens W, Postma DS, Kerstjens HA, Gosens R. Activation of WNT/beta-catenin signaling in pulmonary fibroblasts by TGF-beta(1) is increased in chronic obstructive pulmonary disease. PLoS One 2011;6:e25450.