HemoFLUOR™ PCA

Fluorescence Progenitor Cell Assays for
Cellular Therapy 

 

Download the Progenitor Cell Assay Flyer

 

Buy HemoFLUOR™ PCA Assay Kits
 

Hematopoietic Progenitor Cell Replacement for the CFU Assay

 

To learn how to setup and use HemoFLUOR™ PCA, click on the video links below:

How to Perform a Progenitor Cell Assay (PCA) - An Alternative to All Methylcellulose CFU Assays
 

  • A fluorescence, CFU-equivalent assay for cell processing laboratories.
  • No colony counting required.
  • Transforms the subjective CFU assay into an instrument-based, quantitative assay.
  • Incorporates the same growth factor cocktails used in MethoCult® reagents for direct assay-to-assay comparisons and correlation.
  • Non-subjective, instrument-based and quantitative.
  • Results in 7 days.
  • Faster, easier and more cost effective to use than any traditional CFU assay, even using automated colony counting.
  • Complete assay kit; includes everything needed to culture and obtain results.
  • Permitted by FDA and AABB.

Principle of Single Step Fluorescence Using Resazurin
 

The table below shows the HemoFLUOR™ PCA assays that can replace any methlcellulose, colony-forming unit (CFU) reagent from any competitor. PLEASE NOTE that HemoFLUOR™ PCA incorporates Suspension Expansion Culture™ (SEC™) Technology to replace dispensing error-prone methylcellulose with reagents that use normal pipettes.
 

 
HemoFLUOR™ PCA Number HemoFLUOR™ PCA
Cell Population
Equivalent CFU Cell Population Equivalent MethoCult™
Reagent
Growth Factor Cocktail
PCA1 SC-GEM 2 CFC-GEM 2 H4434 "Classic" EPO, GM-CSF, IL-3, SCF
PCA2 SC-GEM 3 CFC-GEM 3 H4034 "Optimum" EPO, GM-CSF, G-CSF, IL-3, SCF
PCA3 P-GM 1 GM-CFC 1 H4534 "Classic" GM-CSF, IL-3, SCF
PCA4 P-GM 2 GM-CFC 2 H4035 "Optimum" GM-CSF, G-CSF, IL-3, SCF
PCA5 SC-GEMM 3 CFC-GEMM 3 H4435 "Enriched" EPO, GM-CSF, G-CSF, IL-3, IL-6, SCF TPO(#)
PCA6 SC-GEMM CFC-GEMM Methocult "Express" Recombinant cytokines EPO

(#) denotes that the Preferred Cell Systems™ formulation of this product includes TPO that stimulates the production of megakaryopoiesis. MethoCult™ and other competitor formulations of this specific product do not include TPO, do not stimulate the production of megakaryocytes and therefore do not detect the CFC-GEMM stem cell population.

  • Incorporates the measurement of non-fluorescent Resazurin (alimaBlue™) to fluorescent resorufin in an oxidation/reduction reaction that occurs only in live cells. The amount of fluorescence (relative fluorescence units, RFU) is dependent on the metabolic status of the cells and therefore is not only a measure of metabolic viability, but cell proliferation or inhibition.
  • After culture, add 10μL of reagent, mix and measure signal between 30 minutes and 4 hours in a fluorescence plate reader.
  • Uses a plate fluorometer or multimode plate reader to measure fluorescence with an excitation filter 560nm and an emission filter of 590nm.
  • Mobilized peripheral blood
  • Umbilical cord blood
  • Bone marrow
  • Purified cell populations for any of the above tissues
  1. Standardization of the CFU-GM assay using hematopoietic growth factors. J. Hematotherapy:6:191-192 (1997)
  2. Development of a novel assay to evaluate the functional potential of umbilical cord blood progenitors. Transfusion. 48:620-628 (2008).
  3. Potency, Proliferation and Engraftment Potential of Stem Cell Therapeutics: The Relationship between Potency and Clinical Outcome for Hematopoietic Stem Cell Products. J Cell Sci Therapy. (2013).
  4. Detecting primitive hematopoietic stem cells in total nucleated and mononuclear cell fractions from umbilical cord blood segments and units. J Translational Medicine 13:94 (2015)
  5. Improving quality and potency testing for umbilical cord blood: A New Perspective. Stem Cells Translational Medicine. 4:967-973 (2015)
  6. Hematopoietic stem cell potency for cellular therapeutic transplantation. In: Hematopoietic Stem Cells, Ed. Rosana P. Camacho. Published by: InTech Open Access Publisher. ISBN 978-953-307-746-8. (2011).
  7. Measuring the Potency of a Stem Cell Therapeutic. In: Stem Cell Protocols. Methods in Molecular Biology, 1235, Ed. Rich IN. Published by Human Press (2015).
  8. Bioluminescence Potency Measurement of Cellular Therapy Products. In: Cellular Therapy: Principles, Methods, and Regulations, 2nd Edition, Eds. Areman EM and Loper K. Published by AABB.
  9. Present Cord Blood Testing Fails to Determine if the Stem Cells Used for Transplantation are of High Quality and PotencyThe Parents Guide to Cord Blood Foundation
  10. The Difference Between Stem Cell Viability and Potency: A Short Guide for Parent and Patients. The Parents Guide to Cord Blood Foundation

For Research Use Only. Not for clinical diagnostic use.
A fluorescence plate reader or multiparameter plate reader with an excitation filter at 560nm and an emission filter at 590nm.

  • HemoFLUOR™ PCA Master Mix
  • Resazurin reagent
  • 3% SDS solution to stop the reaction
  • Resorufin to standardize the assay
  • Non-sterile, 96-well plate for assay standardization
  • Sterile, 96-well plate(s)
  • Sterile, adhesive foil covers so that unused wells remain sterile

HemoFLUOR™ PCA Assay Manual

QuickGuide for Using HemoFLUOR™ PCA, the instrument-based CFU equivalent progenitor cell assay

When ordering HemoFLUOR™ PCA, simply click on Catalog Number in the table below and you will be taken directly to the desired product.

Cell Population Catalog No. Format MethoCult™ Equivalent
SC-GEM 2 K5-PCA1-1 Low serum H4434 "Classic"
SC-GEM 3 K5-PCA2-1 Low serum H4034 "Optimal"
P-GM 1 K5-PCA3-1 Low serum H4534 "Classic"
P-GM 2 K5-PCA4-1 Low serum H4035 "Optimal"
SC-GEMM 3 K5-PCA5-1 Low serum H4435 "Enriched"
SC-GEMM K5-PCA6-1 Low serum MethoCult "Express"
SC-GEM 2 K5SF-PCA1-1 Serum-Free H4434 "Classic"
SC-GEM 3 K5SF-PCA2-1 Serum-Free H4034 "Optimal"
P-GM 1 K5SF-PCA3-1 Serum-Free H4534 "Classic"
P-GM 2 K5SF-PCA4-1 Serum-Free H4035 "Optimal"
SC-GEMM 3 K5SF-PCA5-1 Serum-Free H4435 "Enriched"
SC-GEMM K5SF-PCA6-1 Serum-Free Metocult "Express"