ColonyGro™
Methylcellulose, Colony-Forming Cell (CFC) or Unit (CFU)
Reagents for Hematopoietic Stem and Progenitor Cells
Uses
- All applications involving lympho-hematopoietic cells with a manual or image analysis readout.
- Clonal growth of leukemic cells.
- 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.
- Superior performance compared to competitor reagents.
- Colonies can be counted after 9-10 days.
- Setup 44 samples/100mL with ColonyGro™, instead of only 33 with reagents from other suppliers.
- Available to detect 8 different stem cell populations, and 10 progenitor and precursor cell populations.
- Unlike competitor formulations, ColonyGro™ CFC-GEMM media contains TPO, allowing the production of erythroid, granulocyte, macrophage and megakaryocytes to be produced as defined for the CFC-GEMM populations.
- Same or lower price than MethoCult™ and other competitor reagents.
- Human
- Non-human primate (Macaca fascicularis (Cynomologus) and Macaca mulatta (Rhesus)
- Dog
- Rat
- Mouse
The following cell populations are available for ColonyGro™. Those populations that have multiple growth factors will produce different colony types in the same culture. Please note that the number of colonies counted in culture containing multiple growth factors will be different to those with single growth factors, which only detect cells in a specific cell compartment.
| Cell Type | Cell Population Detected (1) | MethoCult™ Equivalent | Growth Factor Cocktail | Number of Samples(2) |
|---|---|---|---|---|
| Any | None | NE | None | 44 |
| Stem cell | HPP-SP 1, primitive stem cells | NE | 3,6,SCF,Flt3-L | 44 |
| Stem cell | HPP-SP 2, and all lineages | NE | EPO,GM,2,3,6,7,SCF,TPO, Flt3-L | 44 |
| Stem cell | CFC-GEMM 1, and all hematopoietic lineages | NE | EPO,GM,3,6,SCF,TPO, Flt3-L | 44 |
| Stem cell | CFC-GEMM 2, and all hematopoietic lineages | NE | EPO,GM,3,6,SCF,TPO | 44 |
| Stem cell | CFC-GEMM 3, and all hematopoietic lineages | H4435 "Enriched" MethoCult Express |
EPO,GM,G,3,6,SCF TPO(3) | 44 |
| Stem cell | CFC-GEM 1, and E and GM lineages only(4) | NE | EPO,GM3,6,SCF | 44 |
| Stem cell | CFC-GEM 2, and E and GM lineages only | H4034 "Optimum" | EPO,GM,G,3,SCF | 44 |
| Stem cell | CFC-GEM 3, and E and GM lineages only | H4434 "Classic" | EPO,GM,3,SCF | 44 |
| Progenitor | BFU-E 1, primitive E cells | NE | EPO,3,SCF | 44 |
| Progenitor | BFU-E 2, E cells only | H4430 | EPO (high dose) | 44 |
| Precursor | CFU-E, mature E cells | NE | EPO (low dose) | 44 |
| Progenitor | GM-CFC 1, primitive GM cells | H4534 "Classic" | GM,3,SCF | 44 |
| Progenitor | GM-CFC 2, primitive GM and neutrophils | H4035 "Optimum" | GM,G,3,SCF | 44 |
| Progenitor | GM-CFC 3, GM cells only | NE | GM alone | 44 |
| Precursor | G-CFC, neutrophils only | NE | G alone | 44 |
| Precursor | M-CFC, macrophages only | NE | M alone | 44 |
| Progenitor | Mk-CFC 1, primitive megakryocytes | NE | TPO,3,SCF | 44 |
| Progenitor | Mk-CFC 2, megakaryocytes only | NE | TPO alone | 44 |
| Progenitor | B-CFC, B-lymphocytes | NE | 7 alone | 44 |
NE = No equivalent product available.
(1) = Accepted nomenclature of cell populations detected by the colony-forming cell assay.
(2) = Number of samples that can be performed from 100mL by setting up a total volume of 2.5mL, including 10% cell volume), sufficient for duplicate 1mL cultures in 35mm Petri dishes.
(3) = Contains TPO to stimulate megakaryopoiesis. The equivalent MethoCult™ product does not contain TPO, and therefore cannot be designated as a CFC-GEMM stem cell population, but rather a CFC-GEM population.
(4) = E for erythropoietic; GM for granulocyte-macrophage.
Click image for larger view
The graph to the left shows a cell dose response comparison between ColonyGro™ for the primitive hematopoietic stem cell population, CFC-GEMM 1, and the nearest MethoCult™ equivalent population detected with H4435 media. The difference between the growth factors is given in the table above, but whereas the CFC-GEMM 1 growth factor cocktail contains Flt3-Ligand and thrombopoietin (TPO), to induce the production of megakaryocytes, MethoCult™ H4435, contains neither of these components. Consequently, MethoCult™ H4435 cannot produce megakaryocytes, and therefore does not stimulate the production of primitive stem cells and cannot be designated as a CFC-GEMM population; MethoCult™ H4435 actually stimulates a more mature stem cell population.
- 100mL of ColonyGro™ Reagent
- Sufficient for 44 duplicate samples
- 2.5ml total volume prepared/sample
- 0.25ml sample cell suspension volume required
- 1ml plated in 2 x 35mm Petri dishes
ColonyGro™ is a slightly different formulation that (a) allows it to be easier and more accurately dispensed, (b) allows more samples to be assayed, (c) allows greater cell interaction that is important for cell growth and, (d) allows more rapid growth so that colonies can be detected between 10-12 days instead of 14 days
Do ColonyGro™ reagents contain the same growth factor cocktails as other methylcellulose reagents?
Yes. In some cases, ColonyGro™ actually include more growth factors. For example, the formulation for CFC-GEMM 1 includes thrombopoietin (TPO) to stimulation the production of megakaryocytes. This is not the case for other methylcellulose CFC-GEMM culture reagents produced by other companies who falsely claim that CFC-GEMM can be detected. If TPO is not included, no megakaryocytes will be produced. What is being detected are colonies that contain granulocytes, erythroid cells and macrophages.
How many samples can be used with ColonyGro™?
One of the many advantages of ColonyGro™ is that from a 100mL bottle, 44 samples can be assays as opposed to about 30 for other methylcellulose reagents. The different formulation of methylcellulose combined with the lower total culture volume for duplicate replicates, allows more samples to be assayed. This in turn means that the user can run more samples for about the same price as other methylcellulose reagents.
How many cell populations can be detected using ColonyGro™?
Depending on the species, ColonyGro™ is available to detect up to 19 different cell populations.
What species can be used with ColonyGro™?
Human, non-human primate, dog, rat mouse, horse, sheep and pig.
Why does Preferred Cell Systems recommend using a positive displacement pipette to dispense methylcellulose?
Methylcellulose is notoriously difficult to dispense because it is so viscous. Using a syringe and needle adds to the inaccuracy of the assay because inaccurate volumes are dispensed. By using a positive displacement pipette, methylcellulose can be dispense much more accurately, resulting in lower variation. However, like all pipettes, a positive displacement pipette must be professionally calibrated to dispense accurately.
Why use ColonyGro™ instead of other methylcellulose reagents?
Besides all the reasons given above, ColonyGro™ is competitively priced, so that more samples can be assayed for a similar price. In short, the investigator is getting more for their money.
Can ColonyGro™ be used in the cell processing laboratory?
Yes. Preferred Cell Systems has developed ColonyGro™ with the exact same growth factor cocktails as found in all MethoCult™ reagents used in the cell processing laboratory.
Can ColonyGro™ be used as a cell proliferation assay?
No. Although cell proliferation is needed to produce colonies, the assay cannot measure cell proliferation. The CFC or CFU assay is a differentiation assay because the colonies are left to grown until the cells comprising the colony can be identified as belonging to a specific lineage. This can only occur when the cells have differentiated.
For Research Use Only. Not for clinical diagnostic use.
Inverted microscope with a magnification of 40x - 100x.
Certificate of Analysis (CoA) for each reagent type will be provided with shipment
Download methylcellulose SDS Sheet
When ordering ColonyGro™, simply click the Catalog Number in the table below and you will be taken directly to the desired product.
| Cell Population | Horse | Pig | Sheep |
|---|---|---|---|
| B-CFC | CG-B-100E | CG-B-100P | CG-B-100O |
| BFU-E 1 | CG-BFU1-100E | CG-BFU1-100P | CG-BFU1-100O |
| BFU-E 2 | CG-BFU2-100E | CG-BFU2-100P | CG-BFU2-100O |
| CFC-GEM 1 | CG-GEM1-100E | CG-GEM1-100P | CG-GEM1-100O |
| CFC-GEM 2 | CG-GEM2-100E | CG-GEM2-100P | CG-GEM2-100O |
| CFC-GEMM 1 | CG-GEMM1-100E | CG-GEMM1-100P | CG-GEMM1-100O |
| CFC-GEMM 2 | CG-GEMM2-100E | CG-GEMM2-100P | CG-GEMM2-100O |
| CFU-E | CG-E-100E | CG-E-100P | CG-E-100O |
| G-CFC | CG-G-100E | CG-G-100P | CG-G-100O |
| GM-CFC 1 | CG-GM1-100E | CG-GM1-100P | CG-GM1-100O |
| GM-CFC 3 | CG-GM3-100E | CG-GM3-100P | CG-GM3-100O |
| HPP-SP 1 | CG-HPP1-100E | CG-HPP1-100P | CG-HPP1-100O |
| HPP-SP 2 | CG-HPP2-100E | CG-HPP2-100P | CG-HPP2-100O |
| M-CFC | CG-M-100E | CG-M-100P | CG-M-100O |
| Mk-CFC 1 | CG-Mk1-100E | CG-Mk1-100P | CG-Mk1-100O |
| None | CG-0-100E | CG-0-100P | CG-0-100O |