< OMIPS Directory
OMIP-040

Optimized gating of human prostate cellular subpopulations

Gervaise H. Henry, Nicolas Loof, Douglas W. Strand Cytometry PART A, Volume 91, Issue 12, 1147–1149 (2017)

PURPOSE: This panel was optimized to quantify the relative frequency of the major cell types present in the human prostate in addition to their sorting for downstream applications. Tissue resident white blood cells (referred to here as leukocytes) are identified by CD45, epithelia are identified by CD326, and stroma are double negative with those markers. Epithelia can be further segregated into basal, luminal, and “other” populations using CD26 and CD271. Fibromuscular stroma can be identified by removing CD31‐positive endothelia from the stroma. This panel can also serve as a backbone for the addition of markers to interrogate subpopulations within these major cell types. The panel has been validated on freshly digested and cryopreserved human prostate cells collected from young organ donors, BPH patients, and prostate cancer patients. Other tissue types have not been tested. Other basal cell markers including CD49f, podoplanin, and CD104 are tested and compared with CD271.

CELL TYPE: Freshly digested and cryopreserved primary prostate cells from adult men

MACHINE: BD FACSAria Fusion SORP

Marker Target Clone Fluorochrome Purpose
Dead cells Human Ghost Dye Red 780 viability
CD45 Human H130 PerCP-Cy5.5 leukocyte
CD326 Human EBA-1 BB515 epithelia
NGF Receptor p75 Human ME20.4 PE basal epithelia
CD26 Human BA5b APC luminal epithelia
CD31 Human WM59 Brilliant Violet 421 endothelia