Regeneration and reuse of transwell-type culture inserts for blood-brain barrier modelling

Abstract

Culture inserts of Transwell type are widely used in laboratories worldwide to model non-cerebral vessels, the blood-brain barrier (BBB), and other blood-tissue barriers, and also for the study of chemotaxis and transmigration. However, the use of inserts can generate plastic waste that has an environmental impact due to their one-off design and massive consumption. Thus, it is important to develop a method that can reduce the utilization of inserts but not affect research efficiency. In this study, we propose using a 1:1 (v:v) mix of 30% hydrogen peroxide and 99% sulfuric acid (“piranha solution’’) to completely remove cell debris and matrix from culture inserts. BBB models with inserts regenerated using piranha solution have barrier properties comparable to those of fresh inserts. We show that piranha solution is an effective reagent and allows for the reuse of Transwell-type inserts for BBB modeling up to 5 times. Therefore, the use of this method greatly reduces the production of laboratory waste and benefits numerous laboratories worldwide.