A Cell-Based Assay to Assess the Tau Protein Uptake by Microglia

Abstract

Source: De Marco, D. et al., Cell-based Assay to Study Antibody-mediated Tau Clearance by Microglia. J. Vis. Exp. (2018)

This video showcases a cell-based assay quantifying Tau uptake by murine microglia using pH-sensitive fluorescent-labeled Tau aggregates, analyzed via fluorescence-activated cell sorting for a quantitative assessment.

Protocol

1. BV-2 Microglia Cells Culture NOTE: Handle BV-2 cells under Biosafety Level 2 containment. The BV-2 cell line produces an enveloped recombinant ecotropic retrovirus (capable of infecting murine cells only); such viruses are known for their in vitro transforming ability and in vivo tumorigenic potential. Culture BV-2 cells in high glucose Dulbecco's modified eagle medium (DMEM) supplemented with 10% fetal …

Materials

BV-2 cells	ICLC Interlab Cell Line Collection	ATL03001
Phosphate Buffered Saline (PBS) (1X)	Gibco	10010-015
Trypsin-EDTA 0.05%	Gibco	25300-054
DMEM 4.5 g/dl glucose	Gibco	41966-029
Fetal Bovine Serum	Gibco	10091-148
Penicillin-Streptomycin (10,000 U/mL)	Gibco	15140122
L-Glutamine 200mM	Lonza	17-605E
EasYFlask	Nunc	156499 / 159910
pHrodo Green STP ester	Life Technologies	P35369
Sodium Bicarbonate pH 8.5 100 mM
DMSO	Sigma	D2650-100ml
PD10 columns	GE Healthcare	17-0851-01
BCA Protein Assay Kit	Thermo Fisher Scientific	23225
Greiner CELLSTAR multiwell culture plates	Greiner	665180
Falcon 96-Well Assay Plates	Falcon	353910
Heparin	Sigma	H3393-50KU
Trypsin-EDTA 0.25%	Sigma	T4049-100ml
BSA	Sigma	A7030-100G
EDTA 0.5M, pH8