ColonyGro™

A Traditional Methylcellulose CFU Assay
for 35mm Petri Dishes

 

ColonyGro™

A Traditional Methylcellulose CFU Assay for 35mm Petri Dishes

Cell Population Species  Formulation Catalog Number Quantity
Stem Cell Populations
HPP-SP 1 Human Low serum CG-HPP1-100H 100mL
HPP-SP 2 Human Low serum CG-HPP2-100H 100mL
CFC-GEMM 1 Human Low serum CG-GEMM1-100H 100mL
CFC-GEMM 2 Human Low serum CG-GEMM2-100H 100mL
CFC-GEMM 3 Human Low serum CG-GEMM3-100H 100mL
CFC-GEM 1 Human Low serum CG-GEM1-100H 100mL
CFC-GEM 2 Human Low serum CG-GEM2-100H 100mL
CFC-GEM 3 Human Low serum CG-GEM3-100H 100mL
Progenitor Cell Populations
BFU-E 1 Human Low serum CG-BFU1-100H 100mL
BFU-E 2 Human Low serum CG-BFU2-100H 100mL
GM-CFC 1 Human Low serum CG-GM1-100H 100mL
GM-CFC 2 Human Low serum CG-GM2-100H 100mL
GM-CFC 3 Human Low serum CG-GM3-100H 100mL
Mk-CFC 1 Human Low serum CG-Mk1-100H 100mL
Mk-CFC 2 Human Low serum CG-Mk2-100H 100mL
Precursor Cell Populations
CFU-E Human Low serum CG-E-100H 100mL
G-CFC Human Low serum CG-G-100H 100mL
M-CFC Human Low serum CG-M-100H 100mL
No Growth Factors Added
Any Human Low serum CG-0-100H 100mL

ColonyGro™

A Traditional Methylcellulose CFU Assay for 35mm Petri Dishes

Cell Population Species  Formulation Catalog Number Quantity
Stem Cell Populations
HPP-SP 2 Mouse Low serum CG-HPP2-100M 100mL
CFC-GEMM 1 Mouse Low serum CG-GEMM1-100M 100mL
CFC-GEMM 2 Mouse Low serum CG-GEMM2-100M 100mL
CFC-GEM 1 Mouse Low serum CG-GEM1-100M 100mL
CFC-GEM 3 Mouse Low serum CG-GEM3-100M 100mL
Progenitor Cell Populations
BFU-E 1 Mouse Low serum CG-BFU1-100M 100mL
BFU-E 2 Mouse Low serum CG-BFU2-100M 100mL
GM-CFC 1 Mouse Low serum CG-GM1-100M 100mL
GM-CFC 3 Mouse Low serum CG-GM3-100M 100mL
Mk-CFC 1 Mouse Low serum CG-Mk1-100M 100mL
Precursor Cell Populations
CFU-E Mouse Low serum CG-E-100M 100mL
G-CFC Mouse Low serum CG-G-100M 100mL
M-CFC Mouse Low serum CG-M-100M 100mL
No Growth Factors Added
Any Mouse Low serum CG-0-100M 100mL

ColonyGro™

A Traditional Methylcellulose CFU Assay for 35mm Petri Dishes

Cell Population Species  Formulation Catalog Number Quantity
Stem Cell Populations
HPP-SP 2 Primate Low serum CG-HPP2-100Pr 100mL
CFC-GEMM 1 Primate Low serum CG-GEMM1-100Pr 100mL
Progenitor Cell Populations
BFU-E 1 Primate Low serum CG-BFU1-100Pr 100mL
GM-CFC 1 Primate Low serum CG-GM1-100Pr 100mL
GM-CFC 3 Primate Low serum CG-GM3-100Pr 100mL
Mk-CFC 1 Primate Low serum CG-Mk1-100Pr 100mL
Precursor Cell Populations
CFU-E Primate Low serum CG-E-100Pr 100mL
G-CFC Primate Low serum CG-G-100Pr 100mL
M-CFC Primate Low serum CG-M-100Pr 100mL
No Growth Factors Added
Any Primate Low serum CG-0-100Pr 100mL

ColonyGro™

A Traditional Methylcellulose CFU Assay for 35mm Petri Dishes

Cell Population Species  Formulation Catalog Number Quantity
Stem Cell Populations
HPP-SP 2 Rat Low serum CG-HPP2-100R 100mL
CFC-GEMM 1 Rat Low serum CG-GEMM1-100R 100mL
Progenitor Cell Populations
BFU-E 1 Rat Low serum CG-BFU1-100R 100mL
GM-CFC 1 Rat Low serum CG-GM1-100R 100mL
GM-CFC 3 Rat Low serum CG-GM3-100R 100mL
Mk-CFC 1 Rat Low serum CG-Mk1-100R 100mL
Precursor Cell Populations
CFU-E Rat Low serum CG-E-100R 100mL
G-CFC Rat Low serum CG-G-100R 100mL
M-CFC Rat Low serum CG-M-100R 100mL
No Growth Factors Added
Any Rat Low serum CG-0-100R 100mL

ColonyGro™

A Traditional Methylcellulose CFU Assay for 35mm Petri Dishes

Cell Population Species  Formulation Catalog Number Quantity
Stem Cell Populations
HPP-SP 2 Dog Low serum CG-HPP2-100C 100mL
CFC-GEMM 1 Dog Low serum CG-GEMM1-100C 100mL
Progenitor Cell Populations
BFU-E 1 Dog Low serum CG-BFU1-100C 100mL
GM-CFC 1 Dog Low serum CG-GM1-100C 100mL
GM-CFC 3 Dog Low serum CG-GM3-100C 100mL
Mk-CFC 1 Dog Low serum CG-Mk1-100C 100mL
Precursor Cell Populations
CFU-E Dog Low serum CG-E-100C 100mL
G-CFC Dog Low serum CG-G-100C 100mL
M-CFC Dog Low serum CG-M-100C 100mL
No Growth Factors Added
Any Dog Low serum CG-0-100C 100mL

ColonyGro™

A Traditional Methylcellulose CFU Assay for 35mm Petri Dishes

Cell Population Species  Formulation Catalog Number Quantity
Stem Cell Populations
HPP-SP 2 Horse Low serum CG-HPP2-100E 100mL
CFC-GEMM 1 Horse Low serum CG-GEMM1-100E 100mL
Progenitor Cell Populations
BFU-E 1 Horse Low serum CG-BFU1-100E 100mL
GM-CFC 1 Horse Low serum CG-GM1-100E 100mL
GM-CFC 3 Horse Low serum CG-GM3-100E 100mL
Mk-CFC 1 Horse Low serum CG-Mk1-100E 100mL
Precursor Cell Populations
CFU-E Horse Low serum CG-E-100E 100mL
G-CFC Horse Low serum CG-G-100E 100mL
M-CFC Horse Low serum CG-M-100E 100mL
No Growth Factors Added
Any Horse Low serum CG-0-100E 100mL

ColonyGro™

A Traditional Methylcellulose CFU Assay for 35mm Petri Dishes

Cell Population Species  Formulation Catalog Number Quantity
Stem Cell Populations
HPP-SP 2 Pig Low serum CG-HPP2-100Po 100mL
CFC-GEMM 1 Pig Low serum CG-GEMM1-100Po 100mL
Progenitor Cell Populations
BFU-E 1 Pig Low serum CG-BFU1-100Po 100mL
GM-CFC 1 Pig Low serum CG-GM1-100Po 100mL
GM-CFC 3 Pig Low serum CG-GM3-100Po 100mL
Mk-CFC 1 Pig Low serum CG-Mk1-100Po 100mL
Precursor Cell Populations
CFU-E Pig Low serum CG-E-100Po 100mL
G-CFC Pig Low serum CG-G-100Po 100mL
M-CFC Pig Low serum CG-M-100Po 100mL
No Growth Factors Added
Any Pig Low serum CG-0-100Po 100mL

ColonyGro™

A Traditional Methylcellulose CFU Assay for 35mm Petri Dishes

Cell Population Species  Formulation Catalog Number Quantity
Stem Cell Populations
HPP-SP 2 Sheep Low serum CG-HPP2-100O 100mL
CFC-GEMM 1 Sheep Low serum CG-GEMM1-100O 100mL
Progenitor Cell Populations
BFU-E 1 Sheep Low serum CG-BFU1-100O 100mL
GM-CFC 1 Sheep Low serum CG-GM1-100O 100mL
GM-CFC 3 Sheep Low serum CG-GM3-100O 100mL
Mk-CFC 1 Sheep Low serum CG-Mk1-100O 100mL
Precursor Cell Populations
CFU-E Sheep Low serum CG-E-100O 100mL
G-CFC Sheep Low serum CG-G-100O 100mL
M-CFC Sheep Low serum CG-M-100O 100mL
No Growth Factors Added
Any Sheep Low serum CG-0-100O 100mL

 

ColonyGro™ CFU Assay

 



 

Methylcellulose, colony-forming cell (CFC)/ unit (CFU) reagents for hematopoietic stem and progenitor cells from The Colony Experts.

  • Detect the differentiation potential and/or ability of hematopoietic stem, progenitor and precursor cells by clonal analysis.
  • Manually identify the origin or specific hematopoietic cell populations in culture, based on their in vitro functionality.
  • All applications involving hematopoietic cells with a manual or image analysis readout.
  • Determine the effects of growth factors and/or cytokines on hematopoietic cell differentiation.
  • Determine the growth and differentiation of diseased hematopoietic cells, e.g. leukemic cells. 
  • Correlate in vivo with in vitro differentiation ability.
  • Evaluate ex vivo differential expansion.
  • Evaluate the differentiation/maturation process in response to drugs, xenobiotics or other agents.  
  • Superior performance compared to competitor reagents (see below). 
  • Colonies can be counted after 9-12 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. 

Methylcellulose, Colony-Forming Cell (CFC) or Unit (CFU) Reagents for Hematopoietic Stem and Progenitor Cells
 
  • Human
  • Non-human primate (Macaca fascicularis (Cynomologus) and Macaca mulatta (Rhesus)
  • Horse (Equine)
  • Pig (Porcine)
  • Sheep (Ovine)
  • Dog (Canine)
  • Rat (Rattus)
  • Mouse (Murine)

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) Species Available(5)
Any None NE None 44 H,Pr,E,P,O,C,R,M
Stem cell HPP-SP 1, primitive stem cells NE 3,6,SCF,Flt3-L 44 H, M
Stem cell HPP-SP 2, and all lineages NE EPO,GM,2,3,6,7,
SCF,TPO, Flt3-L
44 H,Pr,E,P,O,C,R,M
Stem cell CFC-GEMM 1, and all hematopoietic lineages MethoCult "Express" EPO,GM,3,6,SCF,
TPO, Flt3-L
44 H,Pr,E,P,O,C,R,M
Stem cell CFC-GEMM 2, and all hematopoietic lineages NE EPO,GM,3,6,SCF,
TPO
44 H, M
Stem cell CFC-GEMM 3, and all hematopoietic lineages H4435 "Enriched"
MethoCult Express
EPO,GM,G,3,6,
SCF TPO(3)
44 H
Stem cell CFC-GEM 1, and E and GM lineages only(4) NE EPO,GM3,6,SCF 44 H, M
Stem cell CFC-GEM 2, and E and GM lineages only H4034 "Optimum" EPO,GM,G,3,SCF 44 H, M
Stem cell CFC-GEM 3, and E and GM lineages only H4434 "Classic" EPO,GM,3,SCF 44 H
Progenitor BFU-E 1, primitive E cells NE EPO,3,SCF 44 H,Pr,E,P,O,C,R,M
Progenitor BFU-E 2, E cells only H4430 EPO (high dose) 44 H, M
Precursor CFU-E, mature E cells NE EPO (low dose) 44 H,Pr,E,P,O,C,R,M
Progenitor GM-CFC 1, primitive GM cells H4534 "Classic" GM,3,SCF 44 H,Pr,E,P,O,C,R,M
Progenitor GM-CFC 2, primitive GM and neutrophils H4035 "Optimum" GM,G,3,SCF 44 H
Progenitor GM-CFC 3, GM cells only NE GM alone 44 H,Pr,E,P,O,C,R,M
Precursor G-CFC, neutrophils only NE G alone 44 H,Pr,E,P,O,C,R,M
Precursor M-CFC, macrophages only NE M alone 44 H,Pr,E,P,O,C,R,M
Progenitor Mk-CFC 1, primitive megakryocytes NE TPO,3,SCF 44 H,Pr,E,P,O,C,R,M
Progenitor Mk-CFC 2, megakaryocytes only NE TPO alone 44 H, M
Progenitor B-CFC, B-lymphocytes NE 7 alone 44 H,Pr,E,P,O,C,R,M


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.

(5) H = Human, Pr = Primate, E = Equine, P = Porcine, O = Ovine, C = Canine, R = Rattus, M = Murine

Preferred Cell Systems™ provides 4 different colony-forming unit assays in 3 different formats. 

ColonyGro™ is one of the 4 CFU assays available in a 35mm Petri dish format. Below, it is compared to the other 3 CFU assays available from Preferred Cell Systems™.

Characteristics AllColonies™:
Traditional/
Micro/Multi
ColonyGro™ CAMEO™-4 CAMEO™-96
Uses: Basic and stem cell research.
Veterinary research.
Limited cellular
therapy.
Basic and stem cell research.
Veterinary research.
Limited cellular
therapy.
Basic and stem cell research.
Veterinary research.
Limited cellular
therapy.
Basic and stem cell research.
Veterinary research.
Limited cellular
therapy.
Limited
hematotoxicity testing
Assay availability: 100mL/50mL/1kit 100mL 50mL 1 kit
Number of samples: 44/50/Depends
on number of
replicates
required(1)
44 50 Depends
on number of
replicates
required(1)
Type of assay: Methylcellulose 
- Clonal
Methylcellulose - Clonal Methylcellulose - Clonal Methylcellulose - Clonal
Assay format: 35mm Petri dish/
4-well Petri dish/
96-well plate
35mm Petri dish 4-well Petri dish 96-well plate
Cell functionality detected: Differentiation or
cell differentiation AND
proliferation
Differentiation Differentiation Cell differentiation AND
proliferation
Readout: Microscopic or microscopic AND
plate reader
Microscopic Microscopic Microscopic AND
plate reader
Readout endpoint: Colony counts or colony counts WITH ATP bioluminescence Colony counts Colony counts Colony counts WITH ATP bioluminescence
Subjectivity: Subjective/subjective/
non-subjective
Subjective Subjective Subjective AND
non-subjective
Cell types detected: Primitive lympho-
hematopoietic stem
cells
Stem, progenitor and/or
precursor cells
Stem, progenitor and/or
precursor cells
Stem and/or
progenitor cells
Species: Human. Primate. Dog. Rat. Mouse Human. primate. Horse. Pig. Sheep. Dog. rat. Mouse Human. primate. Horse. Pig. Sheep. Dog. rat. Mouse Human. primate. Horse. Pig. Sheep. Dog. rat. Mouse
MethoCult™ equivalent assays available: No Yes Yes Yes
Total culture volume required: 2.5mL/0.6mL/
Depends on replicate
number (2)
2.5mL 0.6mL Depends on replicate
number (2)
Number of replicates/culture: 2/4/As many
as required
2 4 As many as required
Volume/culture: 1mL/0.1mL/0.1mL 1mL 0.1mL 0.1mL
Cell culture
time(3):
6-10 days 2-12 days 2-9 days 4-9 days
Instrument calibration: Only for AllColonies™ Multi(4) No No Yes(4)
Standardized assay (5): Only for AllColonies™ Multi No No Yes
Proficiency testing incorporated into assay(6): Only for AllColonies™ Multi No No Yes
Measurement assurance(7): Only for AllColonies™ Multi No No Yes
Validation(8): Only for AllColonies™ Multi No No Yes

(1) = The number of samples tested depends on the number of replicates/sample performed. For example, if 6 replicates/sample are performed, then 16 samples can be analyzed on a single 96-well plate. These samples, however, do not have to performed on the same day.
(2) = The total culture volume for AllColonies™ Multi and CAMEO™-96 depends on the number of replicates/sample performed. If 4 replicates are required, 0.6mL total volume would be prepared, while for 6 replicates, 0.8mL would be prepared.
(3) = Culture time depends on the the state of fractionation or purification of the source cells, cell population tested and species.
(4) = Instrument calibration is performed using the ATP controls included with the assay kit.
(5) = Standardization implies performing a standard curve. ATP standards are included. ColonyGro™ and CAMEO™-4 or any other methylcellulose colony-forming assay from competitors do not include standards or controls and can not be standardized. "Standardized media" does not make an assay "standardized". 
6 = Since AllColonies™ Multi and CAMEO™-96 include standards and controls that allow the results to be compared to measurement assurance parameters every time an assay is performed, proficiency testing is also performed automatically at the same time as the instrument is calibrated and the assay standardized.
7 = Measurement assurance parameters are included in the Instruction Manual.
8 = Validation parameters are included in the Instruction Manual. Since standards and controls are included with these assays, validation can be performed in-house. ColonyGro™ and CAMEO™-4 as well as all other methylcellulose colony-forming assays from competitors, do not include standards and controls and, therefore, cannot be standardized or validated.

How ColonyGro for CFC-GEMM compares to MethoCult H4435  

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
How is ColonyGro™ different from MethoCult™ or other methylcellulose reagents?
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 includes more growth factors. For example, the formulation for CFC-GEMM 1 includes thrombopoietin (TPO) to stimulate 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 grow 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.

ColonyGro™: Methylcellulose Colony-Forming Unit (CFU) Assays

Download the ColonyGro™ Manual
Certificate of Analysis (CoA) for each reagent type will be provided with shipment
Download methylcellulose SDS Sheet