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.

Human Methylcellulose Serum-Free Base Media (Catalog # HSC002SF)

Serum-free media that 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.

Human Methylcellulose Serum-Free Enriched Media without Epo (Catalog # HSC010SF)

A specially formulated serum-free media supplemented with recombinant human (rh) GM-CSF, rhG-CSF, rhIL-3, rhIL-6, 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.

Mouse Methylcellulose Complete Media without Epo (Catalog # HSC008)

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

The products listed above are 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.