Abstract
In vitro screening and testing of drugs and devices is necessary, but in vitro conditions differ greatly from those found in vivo. These differences can lead to false promises of efficacy, or can hide problems of tissue compatibility. Models with ex vivo tissues can be highly valuable bridges which provide relevant matrices for testing [1–9]. Ex vivo tissue models which are closer both biochemically and biophysically can provide useful feedback in a more time- and cost-efficient manner. Herein we describe an ex vivo corneal model for use in drug delivery testing and corneal infection modeling [10]. The protocol covers the tissue harvesting, sterilization, inoculation, and bacterial load quantification. We envision that the model can be used to study bacterial physiology on metabolizable matrices and to study the direct effects of microbial colonization on the cornea's integrity and clarity.•Devitalized cornea.•Non-submersed conditions.•Contact lens compatible.
Highlights
To prevent fungal growth during the threeday study, fluconazole 21.25 μg/ml was added to the soft agar after autoclaving before pouring into sterile 90 mm Petri dishes
Each pathogen was individually streaked for isolation on Tryptic Soy Agar (TSA) plates from frozen stock cultures and incubated for 16–18 h at 37 °C
A 15 ml culture tube containing 5 ml of sterile Tryptic soy broth (TSB) was inoculated with a single colony of the pathogen, vortexed and placed in an orbital shaker at 150 rpm and 37 °C overnight
Summary
To prevent fungal growth during the threeday study, fluconazole 21.25 μg/ml was added to the soft agar after autoclaving before pouring into sterile 90 mm Petri dishes. A 15 ml culture tube containing 5 ml of sterile TSB was inoculated with a single colony of the pathogen, vortexed and placed in an orbital shaker at 150 rpm and 37 °C overnight. About 0.2–0.5 ml bacterial culture was taken from the overnight tube and added to a 250 ml sterile flat-bottom flask containing 20–30 ml TSB.
Sign-in/Register to view highlights & summary 
Published Version (
Free)
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a call