Macrophage cell therapy for cirrhosis

Scientists at the MRC Centre for Regenerative Medicine and the Scottish National Blood Transfusion Centre in Edinburgh have worked together on GMP-compliant manufacture for a novel approach using autologous phagocytic macrophages as a therapeutic approach to mitigate and reverse liver damage in cirrhosis.

As part of the extensive evaluation, in one part they characterised the resulting cells for release criteria and this included DRAQ7(TM) to establish the cell viability alongside the phenotypic marker panel to define the committed phagocytic macrophages: CD45, CD206, CD14 & 25F9.  In their protocol, cells were incubated with FcR block before addition of antibody cocktails, subsequent washing with PEA buffer and addition of DRAQ7 at 3 µM final concentration.

Flow cytometry was performed on both the FACSCanto II and the MACQuant 10. The antibody-chromophore (BV421, FITC, PE, APC) / DRAQ7 panel was compatible with both analysis platforms.  The emission spectrum of DRAQ7 is such that spillover is limited and only in the APC channel.*

* This spillover and requirement for compensation can be completely abrogated by application of the virtual channel method that DRAQ7 uniquely permits. For example, with the chromophores used above, DRAQ7-positive dual-excited events can be displayed in a bivariate plot such as FL3 (PerCP-Cy5.5) vs FL6 (APC-Cy7) or FL4 (PE-Cy7) vs FL5 (APC) in the case of the FACSCanto II. A full explanation of this can be found at www.biostatus.com/DRAQ7 and has been the subject of a poster at ESCCA's annual conference in September 2017.

WHERE TO BUY

Reference:

Fraser, Alasdair R., et al.
"Development, functional characterization and validation of methodology for GMP-compliant manufacture of phagocytic macrophages: A novel cellular therapeutic for liver cirrhosis."
Cytotherapy 19.9 (2017): 1113-1124.