CAMEO™-4

Methylcellulose Colony-Forming Cell (CFC) Assays in Miniaturized Format

 

Buy CAMEO™-4 Assay Kits

Available as assay kits or
contract research services.

 

 

Applications

• All applications involving lympho-hematopoietic cells.
• Detect clonal ability of cells.
• Evaluate the differentiation status and/or potential of different lympho-hematopoietic cell populations.
• Correlate in vivo with in vitro differentiation.
• Differentiation effects of growth factors and or cytokines.
• Evaluate ex vivo differential expansion.
• Evaluate the effects of drugs, xeonbiotics or other agents on the differentiation/maturation process. (Please note that CAMEO™-4 is not recommended for toxicity testing.)

Advantages to using CAMEO™-4 for Your Research?

• Miniaturized assay format requires 1/10 of the volume of reagents, target cells and other sample requirements than the traditional 35mm Petri dish CFC assay.
• Greater number of replicates (4) improves statistics.
• Smaller growth surface for more accurate and faster colony counting.
• No meniscus; allows evaluation of colonies to the well edge.
• Uses the same reagents as HALO®-96 SPCA or HALO®-96 SPCA animalis, allowing a direct comparison and correlation between HALO® and the CFC assay.
• Complete flexibility; evaluate new growth factors and/or cytokines or choose from 17 different cell populations and 8 different species (see below).

Species

CAMEO™-4 can be obtained for the following species:

 

Cell Populations detected using CAMEO™-4 (see Lympho-Hematopoietic System to view cell populations and growth factor/cytokine cocktails)

The differentiation status or potential can be detected using CAMEO™-4 for the following cell populations:

Stem Cells

• HPP-SP 1: IL-3, IL-6, SCF, TPO, Flt3-L
• HPP-SP 2: EPO, GM-CSF, IL-2, IL-3, IL-6, IL-7, SCF, TPO, Flt3-L
• CFC-GEMM 1: EPO, GM-CSF, IL-3, IL-6, SCF, TPO, Flt3-L
• CFC-GEMM 2: EPO, GM-CSF, IL-3, IL-6, SCF, TPO
• CFC-GEM 1: EPO, GM-CSF, IL-3, IL-6, SCF
• CFC-GEM 2: EPO, GM-CSF, IL-3, SCF
• CFC-GEM 3: EPO, GM-CSF, G-CSF, IL-3, SCF

Progenitor Cells

• BFU-E 1: EPO, IL-3, SCF
• BFU-E 2: EPO
• GM-CFC 1: GM-CSF, IL-3, SCF
• GM-CFC 2: GM-CSF, G-CSF, IL-3, SCF
• GM-CFC 3: GM-CSF
• Mk-CFC 1: TPO, IL-3, SCF
• Mk-CFC 2: TPO

Precursor Cells

• CFU-E: EPO
• G-CFC: G-CSF
• M-CFC: M-CSF

Lymphopoietic cells

• T-CFC: IL-2
• B-CFC: IL-7

No growth factors: Add growth factors/cytokines of choice.

Counting Colonies

To count colonies, an inverted microscope is required. Colonies can be counted in two ways as shown in the diagram (click on the diagram to enlarge it).

1. The colonies on the top row are all counted individually, each as a single colony. This is the normal way of counting colonies. It does not take into account that within each of these colonies, several smaller colonies have grown or aggregated together to form one large colony.
2. The lower row shows how each large colony actually consists of smaller colonies that have grown or aggregated together. By counting each of the smaller colonies or "Proliferation Units" (PU), a better estimation of the number of original cells (CFC-GEMM, BFU-E and GM-CFC) is obtained. Note that only a colony that is spherical and has a single center is derived from a single cell. The colonies shown in the lower row are all derived from multiple cells.

 

Use

For Research Use Only. Not for clinical diagnostic use

 

Major Equipment Required

Inverted microscope with a magnification of 40x - 100x.

 

Kit Contents

• CAMEO™-4 Master Mix
• 50, 4-well plates
• Instruction manual

CAMEO™-4 Instruction Manual

To download the manual please go to the Downloads Page.

Related Assays

• CAMEO™-96
• HALO®-96 HuSPCA
• HALO®-96 SPCA animalis
• HALO®-96 PREP
• HemoFLUOR™-96
• HemoLIGHT™-96

CAMEO™-4 for Stem Cell
Processing Laboratories

Methylcellulose Colony-Forming Unit (CFU) Assays for Stem Cell Processing Laboratories, Incorporating the same Growth Factor/Cytokine Cocktails as MethoCult® Reagents

 

Buy CAMEO™-4 assay kits for stem cell processing labs

 

Available as assay kits or
contract research services.

 

CAMEO-4 Stem Cell Processing Laboratory Applications

For all stem cell laboratory applications where MethoCult® reagents are used.

Use in place of MethoCult®.

 

Why Use CAMEO™-4 in place of MethoCult® in the Stem Cell Processing Laboratory?

• Miniaturized assay format requires 1/10 of the volume of reagents, target cells and other sample requirements than the traditional 35mm Petri dish CFU assay.
• Greater number of replicates (4) improves statistics.
• Smaller growth surface for more accurate and faster colony counting.
• No meniscus; allows evaluation of colonies to the well edge.
• Uses the same growth factor/cytokine cocktails as MethoCult® reagents (see below).
• CAMEO™-4 Master Mixes contain the same reagents as HALO®-96 PCA^EQ, allowing direct comparisons and correlations between HALO® and the CFU assay.

 

Species and Tissues

For human

• Mobilized peripheral blood
• Umbilical cord blood
• Bone marrow
• Purified cell populations from the above tissues

 

CAMEO™-4 Equivalent Assays to MethoCult® Reagents

The table below shows the CAMEO™-4 assays that are equivalent to MethoCult® reagents used in the stem cell processing laboratory. Also included are the HALO® equivalent assays (HALO®-96 PCA^EQ) that can be used in place of both CAMEO™-4 assays and MethoCult® reagents.

CAMEO™-4 and HALO®-96 PCA^EQ Assays that are Equivalent to MethoCult® Reagents

CAMEO™-4	HALO®-96 PCAEQ	MethoCult®	Growth Factor/Cytokine Cocktail
KC-GEM2-50H	K2-PCA1-1,2,4*	H4434	EPO, GM-CSF, IL-3, SCF
KC-GEM3-50H	K2-PCA1-1,2,4*	H4034	EPO, GM-CSF, G-CSF, IL-3, SCF
K2-GM1-50H	K2-PCA3-1,2,4*	H4534	GM-CSF, IL-3, SCF
KC-GM2-50H	K2-PCA4-1,2,4*	H4035	GM-CSF, G-CSF, IL-3, SCF

Counting Colonies

To count colonies, an inverted microscope is required. Colonies derived from human bone marrow (shown in the diagram opposite), mobilized peripheral blood or umbilical cord blood, can be counted in two ways as shown in the diagram (click on the diagram to enlarge it).

1. The colonies on the top row are all counted individually, each as a single colony. This is the "traditional" way of counting colonies. It does not take into account that within each of these colonies, several smaller colonies have grown or aggregated together to form one large colony.
2. The lower row shows how each large colony consists of smaller colonies that have grown or aggregated together. By counting each of the smaller colonies or "Proliferation Units" (PU), a better estimation of the number of original colony-forming cells (CFC-GEMM, BFU-E and GM-CFC) can be obtained. Note that only a colony that is spherical and has a single center is derived from a single cell. The colonies shown in the lower row are all derived from several colony-forming cells resulting in the colony being irregular in shape.

 

Use

For Research Use Only. Not for clinical diagnostic use.

 

Major Equipment Required

Inverted microscope with a magnification of 40x - 100x.

 

Kit Contents

• CAMEO™-4 Master Mix
• 50, 4-well plates
• Instruction manual

CAMEO™-4 Assay Kit Manual 

To download the CAMEO™-4 Assay Kit Manual, please go to the Downloads Page.

 

Related Assays

• HALO®-96 PCA^EQ
• HemoFLUOR™-96 PCA^EQ
• HemoLIGHT™-96 PCA^EQ
• STEMpredict™
• HALO®-96 SPC-QC
• HALO®-96 PQR
• HALO®-96 PMT
• HemoFLUOR™-96 PMT
• HemoLIGHT™-96 PMT
• ImmunoGlo™-96
• ImmunoGl0™-MLC
• ImmunoFLUOR™-96
• ImmunoLIGHT™-96

CAMEO™-96

(formerly HALO®-96 MeC)

Clonal methylcellulose proliferation and differentiation assays

 

Buy CAMEO™-96 assay kits

Available as assay kits and
contract research services.

 

 

CAMEO™-96 Applications

• Detection of proliferation AND differentiation of lympho-hematopoietic cells under clonal conditions in a single assay.
• Standardization of the colony-forming cell (CFC) assay.
• Compare and evaluate proliferation and differentiation status and/or potential of different lympho-hematopoietic cell populations from up to 8 species.
• Correlate in vivo with in vitro proliferation and differentiation.
• Distinguish between growth factors and cytokines that have a proliferation effect and/or a differentiation effect.
• Evaluate ex vivo expansion.

Why Use CAMEO™-96 for Your Research?

• CAMEO™-96 is the only assay in which both proliferation and differentiation can be determined simultaneously, using different readouts, in the same assay.
• The only assay that allows standardization of the CFU/CFC assay using standards and controls.
• The only assay that allows conversion of colony counts into standardized and quantitative ATP concentrations.
• Complete flexibility; evaluate growth factors and/or cytokines or choose from 19 different cell populations and 8 species.
• Incorporate powerful and proven bioluminomics™ technology to measure cell proliferation.

Species

CAMEO™-96 is available for the following species:

• Human
• Non-human primate (Macaca fascicularis (Cynomologus) and Macaca mulatta (Rhesus))
• Horse
• Pig
• Sheep
• Dog
• Rat
• Mouse

Cell Populations Detected using CAMEO™-96 (see Lympho-Hematopoietic System to view cell populations and growth factor/cytokine cocktails)

The proliferation and differentiation status or potential can be detected using CAMEO™-96 for the following cell populations:

Stem Cells

• HPP-SP 1: IL-3, IL-6, SCF, TPO, Flt3-L
• HPP-SP 2: EPO, GM-CSF, IL-2, IL-3, Il-6, IL-7, SCF, TPO, Flt3-L
• CFC-GEMM 1: EPO, GM-CSF, IL-3, IL-6, SCF, TPO, Flt3-L
• CFC-GEMM 2: EPO, GM-CSF, IL-3, IL-6, SCF, TPO
• CFC-GEM 1: EPO, GM-CSF, IL-3, IL-6, SCF
• CFC-GEM 2: EPO, GM-CSF, IL-3, SCF
• CFC-GEM 3: EPO, GM-CSF, G-CSF, IL-3, SCF

Progenitor Cells

• BFU-E 1: EPO, IL-3, SCF
• BFU-E 2: EPO
• GM-CFC 1: GM-CSF, IL-3, SCF
• GM-CFC 2: GM-CSF, G-CSF, IL-3, SCF
• GM-CFC 3: GM-CSF
• Mk-CFC 1: TPO, IL-3, SCF
• Mk-CFC 2: TPO

Precursor Cells

• CFU-E: EPO
• G-CFC: G-CSF
• M-CFC: M-CSF

Lymphopoietic cells

• T-CFC: IL-2
• B-CFC: IL-7

No growth factors: Add growth factors/cytokines of choice.

Correlation between Differentiated Colony Counts and Proliferation and Standardization of the CFU/CFC Assay

As shown for CAMEO™-4 and CAMEO™-4 for Stem Cell Processing Laboratories, it is possible to count colonies using two different methods. When the more demanding, but more accurate method of counting Proliferation Units is employed, the number of Proliferation Units counted correlates, in a cell dose-dependent manner, directly with proliferation of the cells producing the Proliferation Units when measured using an ATP-based bioluminescence assay such as CAMEO™-96. The diagram shows that, when Relative Luminescence Units (RLU), as a measure of intracellular ATP (iATP) concentration is detected on day 7 of culture for human bone marrow cells, there is a correlation with the number of Proliferation Units counted either on day 10 or day 14 of culture. This means that measuring iATP on day 7, predicts the outcome on days 10 or 14.

Another way of viewing this correlation is also shown. In this diagram, the results from CAMEO™-4 correlate directly with CAMEO™-96 as a function of cell dose. This correlation, like the one above, is to be expected. The cells first have to proliferate. This proliferation is detected by CAMEO™-96. The same proliferating cells then differentiate and mature, which is detected using CAMEO™-4. 

These results demonstrate that CAMEO™-96 and CAMEO™-4 are detecting the same cells, but using two different readouts. Therefore, CAMEO™-96 can replace CAMEO™-4.

Another important implication of the correlation between measuring proliferation using ATP bioluminescence and colony counting is that the straight-line graph produced between these two parameters allows the total colony counts to be expressed as ATP concentration equivalent units. This is because the ATP bioluminescence assay is calibrated and standardized against an external ATP standard and controls. By using CAMEO™-96 to count differentiated colonies and measure proliferation in the same assay using the same reagents and under the same conditions, the CFU/CFC assays can, for the very first time, be standardized against a second assay. CAMEO™-96 is the only assay available that can standardize the CFU/CFC assay. 

 

Use

For Research Use Only. Not for clinical diagnostic use.

 

Major Equipment Required

Inverted microscope with a magnification of 40x - 100x.
Plate luminometer or multiprmeter plate reader with "glow" luminescence capability.

 

Kit Contents

CAMEO™-96 Assay Kit Manual 

To download the manual, please go to the Downloads Page.

 

Related Assays

• HALO®-96 HuSPCA
• HALO®-96 SPCA animalis
• HALO®-96 PREP
• HemoFLUOR™-96
• HemoLIGHT™-96
• LumiCLONE™-96