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

Published: March 29, 2024

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 …

Disclosures

The authors have nothing to disclose.

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

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Cite This Article
A Cell-Based Assay to Assess the Tau Protein Uptake by Microglia. J. Vis. Exp. (Pending Publication), e22093, doi: (2024).

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