From the Research
The best method for culturing esophagus tissue involves using a 3D organoid culture system with specialized media containing growth factors, as this approach allows for long-term expansion while maintaining the cellular architecture and differentiation potential of esophageal epithelium 1. To obtain esophageal tissue samples, a biopsy is necessary, and the most effective method for this is using a drill instrument, which can safely deliver submucosal tissue samples from submucosal lesions in the upper gastrointestinal tract, resulting in larger total size biopsies and larger submucosal parts of the biopsies compared to standard forceps 1. The biopsy sample should then be digested using collagenase IV (1-2 mg/ml) for 30-60 minutes at 37°C, after which epithelial cells can be isolated and embedded in Matrigel or a similar extracellular matrix. The cells should be cultured in Advanced DMEM/F12 medium supplemented with N2, B27, N-acetylcysteine (1 mM), EGF (50 ng/ml), Noggin (100 ng/ml), R-spondin (500 ng/ml), Wnt3A (100 ng/ml), and Y-27632 ROCK inhibitor (10 μM), and maintained at 37°C with 5% CO2, changing media every 2-3 days. Some key considerations for culturing esophagus tissue include:
- The use of a 3D organoid culture system to maintain cellular architecture and differentiation potential
- The importance of proper biopsy technique, such as using a drill instrument, to obtain high-quality tissue samples
- The need for specialized media and growth factors to support stem cell maintenance and proliferation
- The potential for air-liquid interface cultures to promote stratification and differentiation similar to native esophageal epithelium. It's also worth noting that, while other methods such as endoscopic jejunal biopsy culture may be effective for assessing jejunal microflora, they may not be directly applicable to culturing esophagus tissue 2.