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).