Development of Mouse Fibroblast Cell Line Expressing Human Tau Protein and Evaluation of Tau-Dependent Cytotoxity

摘要

A cell model has been developed to study the fundamental mechanisms of pathogenesis of Alzheimer’s disease (AD). The model is based on the 3T3-4R-tau cell line, clone 47, derived from the parental cell line NIH-3T3 (mouse fibroblasts) permanently expressing the human tau (4R) protein. Stable expression of the tau protein was confirmed by immunofluorescence assay (IFA) and characterized by Western blotting with monoclonal antibodies. Cytotoxicity of different forms of tau protein was estimated in the Transwell two-chamber coculture system involving 3T3-4R-tau cells and primary mouse hippocampal neurons. The data on the toxic effect of human tau protein expressed by 3T3 cells on the primary mouse neurons may be an indirect evidence of a similar scenario of pathological process development in human brain. The 3T3-4R-tau cell line makes it possible to simulate the conditions in human brain during AD and other neurodegenerative diseases more exactly than other cell models and can be used for the directed search of drugs inhibiting neurodegenerative processes.