Fluorescence Based Transwell Macrophage Attraction Assay: An In Vitro Method to Study Tumor-Macrophage Interaction Through Chemical-Induced Chemotaxis

Published: April 30, 2023

Abstract

Source: An, Z. et al. In Vitro Assay to Study Tumor-macrophage Interaction. J. Vis. Exp. (2019)

In this video, we study tumor-macrophage interaction using fluorescence-based transwell macrophage interaction assay. The assay involves the analysis of the migration of macrophages through chemical-induced chemotaxis.

Protocol

1. In Vitro Macrophage Attraction Assay Preparation of Conditioned Media Warm up the tumor cell maintenance medium at a 37 °C water bath for 20 min. Spray the tissue culture hood surface with 70% ethanol and wipe off 70% ethanol on the surface with paper towels. Take the cell detachment solution from the fridge. Spray the bottles of the medium and the cell detachment solution with 70% ethanol, wipe off 70% ethanol on the surface with p…

開示

The authors have nothing to disclose.

Materials

10 mL serological pipettes Olympus plastics 12-104
15 mL sterile centrifuge tubes Olympus plastics 28-103
1 mL pipette tip ART molecular bioproducts 2779-RI
2 mL aspirating pipet Falcon 357558
24-well plate Millipore ECM507 Part of ECM507, or can be
purchased separately
4x lysis buffer Millipore ECM507 Part of ECM507, or can be
purchased separately
5 μm Transwell insert Millipore ECM507 Part of ECM507, or can be
purchased separately
Accutase Innovative cell technologies AT-104
B27 Gibco 12587-010
CyQuant Dye Millipore ECM507 Part of ECM507, or can be
purchased separately
DMEM Gibco 11965-092
DMEM:F12 Gibco 10565-018
EGF Peprotech AF-100-15
FBS Gibco 26140
FGF Peprotech 100-18B
IMDM Gibco 12440-053
PBS Gibco 14190-144
Pen Strep Gibco 15140-122
Trypan blue Biorad 1450021

タグ

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記事を引用
Fluorescence Based Transwell Macrophage Attraction Assay: An In Vitro Method to Study Tumor-Macrophage Interaction Through Chemical-Induced Chemotaxis. J. Vis. Exp. (Pending Publication), e20628, doi: (2023).

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