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CellXVivo Human M2 Macrophage Differentiation Kit

For the differentiation of human CD14+ monocytes into M2 macrophages.
Phenotypic Analysis of Human M2 Macrophages. 
(4)
Differentiated Human M2 Macrophages Secrete IL-10. 
Optimized Base Media in CellXVivo™ Macrophage Differentiation Kits Results in More Efficient Differentiation of M1 and M2 Macrophages. 
CellXVivo Human M2 Macrophage Differentiation Kit
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CDK013

Assay Procedure

Refer to the product datasheet for complete product details.

Briefly, M2 macrophages can be generated from human monocytes using the following procedure:

  • Isolate human CD14+ monocytes from a PBMC preparation
  • Culture CD14+ monocytes in M2 Macrophage Differentiation Media
  • Verify differentiation into M2 macrophages by flow cytometry
  • Verify differentiation into by ELISA detection of IL-10
 

 

Reagents Provided

Reagents Supplied in the CellXVivo™ Human M2 Macrophage Differentiation Kit (Catalog # CDK013):

  • Serum-Free Base Media
  • Recombinant Human M-CSF
  • Reconstitution Buffer 2

 

Other Supplies Required

Reagents

  • MagCellect™ Human CD14+ Cell Isolation Kit (R&D Systems, Catalog # MAGH105, or equivalent).
  • Ficoll-Hypaque™
  • Penicillin (optional)
  • Streptomycin (optional)
  • Cell Dissociation Solution Non-enzymatic 1x (Sigma)
  • Lipopolysaccaride (LPS) (optional)

Equipment

  • Tissue culture flasks and/or plates
  • Inverted microscope
  • Hemocytometer
  • 37 °C and 5% CO2 humidified cell culture incubator
  • Centrifuge
  • Pipettes and pipette tips

 

Procedure Overview

Isolate PBMCs from human blood.

Isolate PBMCs from human blood

Enrich human CD14+ monocytes from PBMCs (e.g., using magnetic cell selection).

Isolate T Cells from PBMCs

Perform a cell count.

Perform a cell count

Suspend 1 x 106 cells/mL in Human M2 Macrophage Differentiation Media.

Culture the cells on plates for 6 days.

Add fresh Human M2 Macrophage Differentiation Media on day 3.

Protocol for Human M2 Macrophage Differentiation Step 5

Verify M2 macrophage differentiation on day 6 by analyzing cell surface marker expression via flow cytometry.

M2 macrophages are ready for downstream application.

Verify Th2 cell differentiation

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FAQs

  • Has the CellXVivo Human M2 Macrophage differentiation kit (Catalog # CDK013) been tested with human monocytic cell lines?

    We have only tested the CellXVivo Human M2 Macrophage differentiation kit with primary CD14+ monocytes derived from PBMCs.

Product Datasheets

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot number in the search box below.

Note: Certificate of Analysis not available for kit components.

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