MLII cell culture

2D in vitro cell culture

  • Airway smooth muscle cells, lung fibroblasts, airway epithelial cells, mast cells
  • All from human origin
  • Analysis of anti-inflammatory efficacy and anti-remodelling efficacy

We use human airway smooth muscle cells, lung fibroblasts, 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;

Anti-inflammatory efficacy

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).
Anti-remodelling efficacy

Inhibition of growth factor (TGF-β, PDGF, WNT, others) induced responses, including:

  • Cell proliferation.
  • 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

 

Previous publications

–        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.

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