Articles

CAMEO-4

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

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

- 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 Research, 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:

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

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-GEMM3: EPO, GM-CSF, G-CSF, IL-3, IL-6, SCF + TPO (Human only. Equivalent to MethoCult® H4435 "Enriched" with TPO added)
- CFC-GEM 1: EPO, GM-CSF, IL-3, IL-6, SCF
- CFC-GEM 2: EPO, GM-CSF, IL-3, SCF (Equivalent to MethoCult® H4434 "Classic")
- CFC-GEM 3: EPO, GM-CSF, G-CSF, IL-3, SCF (Human only. Equivalent to MethoCult® H4034 "Optimum")

Progenitor Cells

- BFU-E 1: EPO, IL-3, SCF
- BFU-E 2: EPO
- GM-CFC 1: GM-CSF, IL-3, SCF (Equivalent to MethoCult® H4534 "Classic")
- GM-CFC 2: GM-CSF, G-CSF, IL-3, SCF (Human only. Equivalent to MethoCult® H4035 "Optimum")
- 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