Human Methylcellulose Serum-Free Base Media

Defined media for human hematopoietic stem cell growth and differentiation.

Why use R&D Systems Human Methylcellulose Serum-Free Base Media for Colony Forming Cell Assays?

Colony forming cell (CFC) assays, which are used to enumerate and quantify multi-potent and single lineage hematopoietic progenitors, can be time consuming and laborious.

Successful growth and enumeration of cell colonies is dependent on factors such as accurate cell counts, the presence of growth factors and/or cytokines, adequate humidity, and the use of high quality media. Additionally, inter-assay variation can be decreased by using defined media. R&D Systems offers Human Methylcellulose Serum-Free Base Media with superior optical clarity to support optimal colony growth, enumeration, and identification.

R&D Systems Human Methylcellulose Serum-Free Base Media:

• Optical clarity facilitates colony identification.
• High lot-to-lot consistency decreases variation.
• Supports reproducible in vitro growth of hematopoietic stem and progenitor cells.
• Can be supplemented with user-defined cytokines and growth factors.
• Increased cloning efficiency and improved colony growth compared to agar.

• 90 mL of Human Methylcellulose Serum-Free Base Media and 15 mL of Cell Resuspension Solution.

Stability and Storage

Human Methylcellulose Base Media and the Cell Resuspension Solution should be stored at ≤-20 °C upon receipt. Storage at 2 °C to 8 °C is not recommended.

Precautions

• The acute and chronic effects of overexposure to this media are unknown. Safe laboratory procedures should be followed and protective clothing should be worn when handling this media.
• The human Transferrin used in this product was derived from human plasma, which has been tested and found negative for HIV-1/2 antibodies, Hepatitis B surface antigen, Hepatitis C antibody, Syphilis, and p24 antigen by FDA approved methods. Handle as if capable of transmitting infection, and dispose of according to applicable regulations.

Limitations

• The safety and efficacy of this product in diagnostic or other clinical uses has not been established.
• The reagent should not be used beyond the expiration date indicated on the vial labels.
• The media is optimized to assay human hematopoietic progenitors and is ineffective with mouse hematopoietic progenitors.
• Derivation of human hematopoietic progenitors from different individuals may cause results to vary.

Human Hematopoietic Colony Formation Using the Methylcellulose-based Colony Forming Cell Assay. A. Colony forming unit-erythroid (CFU-E) are 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. B. Colony forming unit-granulocyte (CFU-G) are clonogenic progenitors of granulocytes that give rise to a homogeneous population of eosinophils, basophils, or neutrophils. C. Colony forming unit-granulocyte, macrophage (CFU-GM) are 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. D. Burst forming unit-erythroid (BFU-E) colonies 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. E. Colony forming unit-macrophage (CFU-M) are clonogenic progenitors of macrophages that give rise to a homogenous population of macrophages. F. Colony forming unit-granulocyte, erythrocyte, macrophage, megakaryocyte (CFU-GEMM) are multi-lineage progenitors that give rise to erythroid, granulocyte, macrophage and megakaryocyte lineages, as the name indicates.

Human Methylcellulose Stock and Base Media

Complete Human Methylcellulose Media

*Base media and stock solutions do not contain cytokines and will not support colony growth unless conditioned media, cytokines, or other culture supplements are added.

The definitive hematopoietic system is made up of all adult blood cell types including megakaryocytes, erythrocytes, and cells of the myeloid and lymphoid lineages. All of these cells are derived from multipotent hematopoietic stem cells (HSCs) through a succession of precursors with progressively limited potential. Hematopoietic stem cells are tissue-specific stem cells that exhibit remarkable self-renewal capacity and are responsible for the life-long mainte­nance of the hematopoietic system. HSCs are rare cells that reside in adult bone marrow where hematopoiesis is continu­ously taking place. They can also be found in cord blood, fetal liver, adult spleen, and peripheral blood. R&D Systems offers several products for studying hematopoietic lineage cells including serum-free media, lineage deple­tion antibodies and kits, and reagents for performing colony forming cell (CFC) assays.