4.4 Article

Adapting the Electrospinning Process to Provide Three Unique Environments for a Tri-layered In Vitro Model of the Airway Wall

Journal

JOVE-JOURNAL OF VISUALIZED EXPERIMENTS
Volume -, Issue 101, Pages -

Publisher

JOURNAL OF VISUALIZED EXPERIMENTS
DOI: 10.3791/52986

Keywords

Bioengineering; Issue 101; Electrospinning; 3D Cell Culture; Bioreactor; Airway; Tissue Engineering; In Vitro Model

Funding

  1. European Union [270194]
  2. National Centre for the Replacement, Refinement, and Reduction of Animals in Research (NC3Rs)
  3. Engineering and Physical Research Centre (EPSRC) Doctoral Training Centre (DTC) in Regenerative Medicine, U.K
  4. BBSRC [BB/H011293/1] Funding Source: UKRI
  5. MRC [G1001367] Funding Source: UKRI
  6. Biotechnology and Biological Sciences Research Council [BB/H011293/1] Funding Source: researchfish
  7. Medical Research Council [G1001367] Funding Source: researchfish
  8. National Centre for the Replacement, Refinement and Reduction of Animals in Research (NC3Rs) [G1001804/1] Funding Source: researchfish

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Electrospinning is a highly adaptable method producing porous 3D fibrous scaffolds that can be exploited in in vitro cell culture. Alterations to intrinsic parameters within the process allow a high degree of control over scaffold characteristics including fiber diameter, alignment and porosity. By developing scaffolds with similar dimensions and topographies to organ-or tissue-specific extracellular matrices (ECM), micro-environments representative to those that cells are exposed to in situ can be created. The airway bronchiole wall, comprised of three main micro-environments, was selected as a model tissue. Using decellularized airway ECM as a guide, we electrospun the non-degradable polymer, polyethylene terephthalate (PET), by three different protocols to produce three individual electrospun scaffolds optimized for epithelial, fibroblast or smooth muscle cell-culture. Using a commercially available bioreactor system, we stably co-cultured the three cell-types to provide an in vitro model of the airway wall over an extended time period. This model highlights the potential for such methods being employed in in vitro diagnostic studies investigating important inter-cellular crosstalk mechanisms or assessing novel pharmaceutical targets, by providing a relevant platform to allow the culture of fully differentiated adult cells within 3D, tissue-specific environments.

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