Methylcellulose-based Reagents for Colony Forming Cell Assays

• Procedure for the Human Colony Forming Cell (CFC) Assay Using Methylcellulose-based Media
• Procedure for the Mouse Colony Forming Cell (CFC) Assay Using Methylcellulose-based Media

Methylcellulose Stock Solution (Catalog # HSC001)

Contains 3% methylcellulose in Iscove's MDM and can be used in both human and mouse hematopoietic stem cell research.

Human Methylcellulose Base Media (Catalog # HSC002)

Contains all of the basic components with the exception of the cytokines required to perform human CFC assays for hematopoietic stem cell research.

Mouse Methylcellulose Base Media (Catalog # HSC006)

Contains all of the basic components with the exception of the cytokines required to perform mouse CFC assays for hematopoietic stem cell research.

Human Methylcellulose Complete Media (Catalog # HSC003)

A specially formulated media supplemented with recombinant human (rh) GM-CSF, rhIL-3, rhSCF, and rhEpo optimized for CFC assays of mouse clonogenic hematopoietic progenitors from human bone marrow, peripheral blood, cord blood, leukopheresis products, and purified CD34⁺ cells.

Mouse Methylcellulose Complete Media (Catalog # HSC007)

A specially formulated media supplemented with recombinant mouse (rm) IL-3, rmIL-6, rmSCF, and recombinant human Epo optimized for CFC assays of mouse clonogenic hematopoietic progenitors from mouse bone marrow, peripheral blood, spleen, and fetal liver.

Human Methylcellulose Enriched Media (Catalog # HSC005)

A specially formulated media supplemented with recombinant human (rh) GM-CSF, rhG-CSF, rhIL-3, rhIL-6, rhSCF, and rhEpo optimized for use with purified CD34⁺ cells in the CFC assay at the end of the long-term culture-initiating cell (LTC-IC) assay.

Human Methylcellulose Serum-Free Enriched Media (Catalog # HSC005SF)

A specially formulated serum-free media supplemented with recombinant human (rh) GM-CSF, rhG-CSF, rhIL-3, rhIL-6, rhSCF, and rhEpo optimized for CFC assays of human clonogenic hematopoietic progenitors from human bone marrow, perihperal blood, cord blood, leukopheresis products, and purified CD34⁺ cells.

The products listed above are for the enumeration of:

• CFU-E (colony-forming unit-erythroid)
• BFU-E (burst-forming unit-erythroid)
• CFU-G (colony-forming unit-granulocyte)
• CFU-M (colony-forming unit-macrophage)
• CFU-GM (colony-forming unit-granulocyte/macrophage)
• CFU-GEMM (colony-forming unit-granulocyte/erythroid/macrophage/megakaryocyte)

Human Methylcellulose Complete Media without Epo (Catalog # HSC004)

A specially formulated media supplemented with recombinant human (rh) GM-CSF, rhIL-3, and rhSCF optimized for CFC assays of human clonogenic hematopoietic progenitors from human bone marrow, peripheral blood, cord blood, leukopheresis products, and purified CD34⁺ cells.

The product listed above is for the enumeration of:

• CFU-GM
• CFU-G
• CFU-M

Examples of colony types

CFU-E (Colony forming unit-erythroid): Clonogenic progenitors that produce only one or two clusters with each cluster containing from 8 to approximately 100 hemoglobinized erythroblasts. It represents the more mature erythroid progenitors that have less proliferative capacity.	CFU-G (Colony forming unit-granulocyte): Clonogenic progenitors of granulocytes that give rise to a homogeneous population of eosinophils, basophils or neu­trophils.	CFU-GM (Colony forming unit-granulocyte, macrophage): Progenitors that give rise to colonies containing a heterogene­ous population of macrophages and granulocytes. The mor­phology is similar to the CFU-M and CFU-G descriptions.
BFU-E (Burst forming unit-erythroid): The size of the colony can be described as small (3 to 8 clusters), intermediate (9 to 16 clusters), or large (more than 16 clusters) according to the number of clusters present. These are primitive erythroid progenitors that have high proliferative capacity.	CFU-M (Colony forming unit-macrophage): Clonogenic progenitors of macrophages that give rise to a ho­mogenous population of macrophages.	CFU-GEMM (Colony forming unit-granulocyte, erythrocyte,macrophage, megakaryocyte): Multi-lineage progenitors that give rise to erythroid, granulo­cyte, macrophage and megakaryocyte lineages, as the name indicates.