CAMEO™-96

Complete, Standardized, Methylcellulose Colony-Forming Cell (CFC) or Unit (CFU)
Proliferation and Differentiation Assay for Hematopoietic Stem and Progenitor Cells

 

Buy CAMEO™-96 Assay Kits
 

Complete, standardized, methylcellulose colony-forming cell (CFC) / unit (CFU) proliferation and differentiation assay for hematopoietic stem and progenitor cells from The Colony Experts.

 

  • All applications involving lympho-hematopoietic cells with a manual or image analysis readout.
  • Leukemic cell proliferation and differentiation.
  • Detect clonal ability of cells.
  • Evaluate the differentiation status or potential of different lympho-hematopoietic cell populations.
  • Correlate in vivo with in vitro differentiation.
  • Differentiation effects of growth factors or cytokines.
  • Evaluate ex vivo differential expansion.
  • Evaluate the effects of drugs, xenobiotics or other agents on the differentiation/maturation process.  
  • CAMEO™-96 is the only complete assay in which both proliferation and differentiation can be determined simultaneously, using different readouts.
  • Allows standardization of the CFU/CFC assay using standards and controls.
  • Allows conversion of colony counts into standardized and quantitative ATP concentrations.
  • Complete flexibility; evaluate growth factors or cytokines.
  • Available for 19 different cell populations and 8 species.
  • Incorporates powerful ATP bioluminescence readout technology to measure cell proliferation.

Complete, Standardized, Methylcellulose Colony-Forming Cell (CFC) or Unit (CFU) Proliferation and Differentiation Assay for Hematopoietic Stem and Progenitor Cells
 
All ATP bioluminescence assays kits from Preferred Cell Systems™ include an ATP standard(s) and controls.  Performing the ATP standard curve and controls is highly recommended. The controls are used to calibrate the luminescence plate reader and the ATP standard is used to standardize the assay. The values obtained from the standard curve and controls (see diagram below) are then compared to the measurement assurance parameters that are provided in the instruction manual. If the values obtained are within the ranges provided by the measurement assurance parameters, you, the user, have successfully performed the proficiency testing to ensure that you have not only performed the assay correctly, but that the results you obtain can be trusted.

No additional proficiency testing is required if the calibration and standardization procedure is performed. The values you obtain from each calibration and standardization can be logged and used for certification that the assay has been performed correctly and that the results are trustworthy.

Calibration and standardization of an ATP bioluminescence assay from Preferred Cell Systems™ - Measurement Assurance
 
  • Human
  • Non-human primate (Macaca fascicularis (Cynomologus) and Macaca mulatta (Rhesus))
  • Horse
  • Pig
  • Sheep
  • Dog
  • Rat
  • Mouse

The following cell populations are available as CAMEO™-96 Complete Assay Kits.  Those populations that have multiple growth factors will produce different colony types in the same culture. Please note that the number of colonies counted and their proliferation in cultures containing multiple growth factors will be different to those with single growth factors, which only stimulate cells in a specific cell compartment.
 

Cell Type Cell Population Detected (1) MethoCult™ Equivalent Growth Factor Cocktail Suggested Number of Samples(2)
Any None NE None 24, 16 or 12
Stem cell HPP-SP 1 NE 3,6,SCF,Flt3-L 24, 16 or 12
Stem cell HPP-SP 2 NE EPO,GM,2,3,6,7,SCF,TPO, Flt3-L 24, 16 or 12
Stem cell CFC-GEMM 1 NE EPO,GM,3,6,SCF,TPO, Flt3-L 24, 16 or 12
Stem cell CFC-GEMM 2 NE EPO,GM,3,6,SCF,TPO 24, 16 or 12
Stem cell CFC-GEM 1 NE EPO,GM3,6,SCF 24, 16 or 12
Stem cell CFC-GEM 2 H4034 EPO,GM,G,3,SCF 24, 16 or 12
Progenitor BFU-E 1 NE EPO,3,SCF 24, 16 or 12
Progenitor GM-CFC 1 H4534 GM,3,SCF 24, 16 or 12
Progenitor Mk-CFC 1 NE TPO,3,SCF 24, 16 or 12
Progenitor T-CFC NE 2 alone 24, 16 or 12
Progenitor B-CFC NE 7 alone 24, 16 or 12


NE = No equivalent product available.

(1) = Accepted nomenclature of cell populations detected by the colony-forming cell assay.

(2) = The number of samples is based on the number of replicate wells/sample. If 4 replicate wells/sample are used, 24 samples/plate can be detected. Using 6 replicate wells/sample, 16 samples can be detected and for 8 replicates wells/samples prepared in columns, 12 samples can be measured.

For Research Use Only. Not for clinical diagnostic use.

Inverted microscope with a magnification of 40x - 100x.
Luminescence plate reader or multimode plate reader with "glow" luminescence capability.
  • CAMEO™-96 Reagent
  • ATP standard
  • ATP high and low controls
  • ATP Enumeration Reagent
  • Sterile, 96-well plates
  • Non-sterile-96-well plates
  • Sterile, adhesive foil covers
What is the difference between CAMEO-™4 and CAMEO™-96?
There are many differences between these 2 assays. CAMEO™-4 is a traditional miniaturized methylcellulose colony-forming assay. CAMEO™-96 is also a methylcellulose colony forming assay, but is performed in a clear bottom, 96-well plate in which colonies can be counted and cell proliferation within the colonies measured using the most sensitive and accurate measurement of cell proliferation available, namely ATP bioluminescence. CAMEO™-96 also distinguishes itself from all other methylcellulose assays it that the colony-forming cell (CFC) assay is fully standardized. Standardization of the CFC assay is otherwise impossible with traditional methylcellulose assay.
 
How is the CAMEO™-96 methylcellulose CFC assay standardized?
After cell culture, the colonies are counted under an inverted microscope. Prior to measuring cell proliferation, this second step of the assay is calibrated and standardized against an ATP standard. The cultures are then processed and the amount of cell proliferation is determined in a luminescence plate reader. The amount of proliferation correlated directly with the concentration of intracellular ATP produced by the cells. When the total number of colonies is plotted against the ATP concentration, the number of colonies can be directly interpolated into standardized ATP concentrations. Since the ATP step of the assay is standardized, the colony counting step is also standardized.
 
How many replicate wells are needed for a CAMEO™-96 assay?
Preferred Cell Systems™ recommends a minimum number of 4 replicate wells per data point. Six replicate wells are better from a statistical viewpoint. However, the investigator is flexible to use any number of replicate wells they choose.
 
What species and cell populations are available with CAMEO™-96?
CAMEO™-96 assay kits are available for human, primate, dog rat and mouse cells. The assay kits are available for up to 11 different populations from each species.
 
How long does CAMEO™-96 take to perform?
This depends on the species. However, in general, CAMEO™-96 for human cells is a 7 day assay, while that for all other species is a 5 day assay. This is much faster than for any traditional CFC assay.
 
How long does the ATP bioluminescence step take?
The calibration and standardization of the assay will take between 20-30min to prepare and read the results. A full plate of samples will take about 15-20min to process, followed by a 10min incubation in the dark and less than 5 min to measure.
 
Is there a non-methylcellulose version of CAMEO™-96?
Yes. The non-methylcellulose version of CAMEO™-96 is called HALO® and incorporate Suspension Expansion Culture™ (SEC™) Technology.
 

CAMEO™-96: The only Standardized Colony-Forming Unit (CFU) Assay for Proliferation and Differentiation of Primitive Hematopoietic Cells

Download the CAMEO™-96 Technical Manual
Download the ATP Optimization Kit Protocol for First-Time Users
Download Luminometer Setup and RLU to ATP Conversion
Download Certificate of Analysis (CoAs) for ATP Bioluminescence Components. Note: Some CoAs will be included with assay kit
Download methylcellulose SDS sheet
Download ATP standards and controls SDS sheet
Download ATP Enumeration Reagents (ATP-ER) SDS sheet