Complete, Standardized, Methylcellulose Colony-Forming Cell (CFC) or Unit (CFU)
Proliferation and Differentiation Assay for Hematopoietic Stem and Progenitor Cells
- 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.
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.
- Non-human primate (Macaca fascicularis (Cynomologus) and Macaca mulatta (Rhesus))
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.
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
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.
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
When ordering CAMEO-96™, simply click the Catalog Number in the table below and you will be taken directly to the desired product.