Mouse Methylcellulose Base Media

Name: Mouse Methylcellulose Base Media
Brand: R&D Systems
SKU: HSC006

R&D Systems, part of Bio-Techne | Catalog # HSC006

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Product Summary
Scientific Data
Preparation & Storage
Background
Product Documents
Specific Notices

Key Product Details

Species

Mouse

Product Summary for Mouse Methylcellulose Base Media

Kit Summary

For the support of mouse hematopoietic stem cell growth and differentiation.

Key Benefits

Can be supplemented with user-defined growth factors
High lot-to-lot consistency decreases variation
Excellent optical clarity for easy identification of colonies

Why use R&D Systems Mouse Methylcellulose 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. R&D Systems offers Mouse Methylcellulose Base Media with superior optical clarity to support optimal colony growth, enumeration, and identification. The Mouse Methylcellulose Base Media contains components that have been optimized for CFC assays. Individual researchers can customize the media by adding cells and other culture supplements tailored to their specific research. This product can also be used in the long-term culture-initiating cell (LTC-IC) assay.

R&D Systems Mouse Methylcellulose Base Media:

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.
Optical clarity facilitates colony identification.
High lot-to-lot consistency decreases variation.

Kit Contents

90 mL of Mouse Methylcellulose Base Media.

Contents	Concentration (when diluted to a final volume of 100 mL)
Methylcellulose (1500 cps) in Iscove’s Modified Dulbecco's Medium	1.4%
Fetal Bovine Serum	15%
Bovine Serum Albumin	2%
L-Glutamine	2 mM
2-Mercaptoethanol	5 x 10^-5 M
Recombinant Human Insulin	10 μg/mL
Human Transferrin	200 μg/mL

Stability and Storage

Mouse Methylcellulose Base Media 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, ALT Test and NAT-PCR (HAV, HIV, HBC, HCV, and Parovirus B19) 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 label.
Derivation of mouse hematopoietic progenitors from different individual animals may cause results to vary.
The media is optimized to assay mouse hematopoietic progenitors and is ineffective with human hematopoietic progenitors.

Guide to Choosing Media for the Colony Forming Cell (CFC) Assay

Mouse Methylcellulose Stock and Base Media

Catalog #	Product Description	Volume	Colonies Selected for	Contains Serum	Cytokines Included
HSC001	Methylcellulose Stock Solution	100 mL	N/A*	No	None
HSC006	Mouse Methylcellulose	90 mL	N/A*	Yes	None
HSC011	StemXVivo^® Methylcellulose Concentrate	50 mL	N/A*	No	None

Complete Mouse Methylcellulose Media

Catalog #	Product Description	Volume	Colonies Selected for	Contains Serum	Cytokines Included
HSC007	Mouse Methylcellulose Complete Media	100 mL	BFU-E CFU-E CFU-G CFU-GEMM CFU-GM CFU-M	Yes	Epo IL-3 IL-6 SCF

*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.

Scientific Data Images for Mouse Methylcellulose Base Media

Mouse Hematopoietic Colony Formation Using the Methylcellulose-based Colony Forming Cell Assay.

Burst forming unit-erythroid (BFU-E) colonies are defined as clusters with a minimal of 30 cells that can be seen from day 7 onward. Each individual cluster consisted of tiny, irregular shaped cells that may appear fused together. Each cluster normally contains 5-8 cells, and the size of the cluster is similar to that of a single macrophage. The cluster may vary in sizes and color. A large BFU-E is usually bright red and is differentiable even without the use of a microscope. Smaller BFU-E may not appear red in color but is distinguishable based on the morphology.B.Colony forming unit-macrophage (CFU-M; left) are clonogenic progenitors of macrophages that give rise to a homogenous population of macrophages. Colony forming unit-granulocyte (CFU-G; right) 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.Colony forming unit-granulocyte, erythrocyte, macrophage, megakaryocyte (CFU-GEMM) are multi-lineage progenitors that give rise to the lineage of erythroid, granulocytes, macrophages, and megakaryocytes as the name indicates. It can be identified as reddish colored cells (erythroid) mixed with colorless cells (granulocytes, macrophages, and megakaryocytes) in a single colony. This progenitor is typically the largest colony on the culture dish; occasionally CFU-GM may attain a size comparable or larger than that of CFU-GEMM.

Formulation, Preparation, and Storage

Shipping

The product is shipped with dry ice or equivalent. Upon receipt, store it immediately at the temperature recommended below.

Storage

Store the unopened product at -20 to -70 °C. Use a manual defrost freezer and avoid repeated freeze-thaw cycles. Do not use past expiration date.

Background: Hematopoietic Stem Cells

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 mainteÂnance of the hematopoietic system. HSCs are rare cells that reside in adult bone marrow where hematopoiesis is continuÂously 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 depleÂtion antibodies and kits, and reagents for performing colony forming cell (CFC) assays.