Semi-intact Cell System and Cell Resealing Technique for Image-based Analysis of Intracellular Events: A Powerful Cellular Tool for Single Cell Biology

Abstract

Semi-intact cells are cells whose plasma membrane is permeabilized with toxins by creating pores. Semi-intact cells allow for the exchange of the original cytosolic proteins with an exogenously-made cytosol prepared from cells of varying states, such as cells at different stages of the cell cycle, differentiation, or cells in a diseased state. Recently we found that the pores in semi-intact cells can be resealed by calcium ions. Here, we introduce a basic protocol for preparing a “disease-model cell” using the resealing technique of semi-intact cells, in which the cytosolic conditions of a diabetic liver was replicated, and demonstrate the disturbances in endocytosis and signal transduction in the diabetic liver-model cells.