StemXVivo Serum-Free Tumorsphere Media
R&D Systems, part of Bio-Techne | Catalog # CCM012
Key Product Details
Summary for StemXVivo Serum-Free Tumorsphere Media
Semi-solid media formulated and optimized for tumorsphere formation.
Key Benefits
- Shown to support sphere formation of seven tumor cell lines
- Lot-to-lot consistency reduces experimental variability
- Specially formulated and optimized for tumorsphere formation
Why Perform the Tumorsphere Assay Using Specially Formulated Media?
Tumorsphere assays, also called sphere-formation assays, are in vitro assays that assess stem cell characteristics of tumor cells. There are a number of variables that contribute to variation in data obtained from tumorsphere assays including the purity of the starting cell population, the type and quantity of media and supplements, and the potential for tumorspheres to aggregate.
To minimize experimental variability due to the media and supplements, R&D Systems offers StemXVivo® Serum-Free Tumorsphere Media, which is formulated and optimized for tumorsphere formation.
StemXVivo® Serum-Free Tumorsphere Media:
- Shown to support sphere formation of seven tumor cell lines.
- Formulated and optimized for efficient tumorsphere formation.
- Lot-to-lot consistency reduces experimental variability.
Kit Contents
Media Components
- 100 mL of StemXVivo® Serum-Free Tumorsphere Media.
Stability and Storage
Upon receipt, StemXVivo® Serum-Free Tumorsphere Media should be stored at =-20 °C in a manual defrost freezer. The media can be thawed at 2 °C to 8 °C or at room temperature. Thawed media can be aliquoted and stored at =-20°C in a manual defrost freezer for up to 3 months or used within two weeks when stored in the dark at 2 °C to 8 °C. Avoid repeated freeze-thaw cycles. Do not use this product beyond the expiration date.
Precautions
This product contains human transferrin. The transferrin was purified from donor plasma and tested at the donor level using an FDA licensed method and found to be non-reactive for anti-HIV-1/2 and Hepatitis B surface antigen.
Limitations
- FOR LABORATORY RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES.
- The safety and efficacy of this product in diagnostic or other clinical uses has not been established.
- This reagent should not be used beyond the expiration date indicated on the label.
- Results may vary with cells cultured by different methods.
As for all stem cells, oncogenically transformed cancer stem cells (CSCs) are defined by their ability for self-renewal and multipotency. The CSC hypothesis states that, although CSCs represent a rare population of cells within a tumor, their high tumorigenic capacity drives tumorigenesis. Due to their intrinsic stem cell-like properties, CSC proliferation generates more CSCs, and all the differentiated cell types that compose the bulk of the tumor. Non-CSCs in the tumor have been shown to proliferate at a faster rate than CSCs, but have little tumor-initiating potential. Because CSCs exhibit increased resistance to toxic and chemical insults, this specific subpopulation of cells is believed to underlie resistance to chemotherapy and disease relapse. In fact, the CSC model posits that all CSCs must be eradicated to eliminate a tumor and prevent its recurrence.
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Tumorspheres Formed from the MCF-7 Human Breast Cancer Cell Line. MCF-7 human breast cancer cells were suspended in StemXVivo®Serum-Free Tumorsphere Media (Catalog # CCM012) containing 2 U/mL Heparin (Tocris Catalog # 2812) and 0.8 µg/mL Hydrocortisone (Tocris, Catalog # 4093). The cells were plated at 3 x 104cells per well in a 6-well ultralow adhesion culture plate and cultured at 37°C and 5% CO2for 3-10 days to induce tumorsphere formation. Tumorspheres were imaged using a CloneSelect™Imager from Molecular Devices (New Milton) Limited. |
Assay Procedure
Refer to the product datasheet for complete product details.
Briefly, tumorsphere formation can be induced using the following procedure
- Cells are suspended in StemXVivo Serum-Free Tumorsphere Media containing Heparin and Hydrocortisone
- Cells are cultured on low adhesion plates at 37 °C and 5% CO2
- Tumorspheres should form in 3-10 days
Reagents Provided
Reagents supplied in the StemXVivo® Serum-Free Tumorsphere Media (Catalog # CCM012):
- 100 mL of StemXVivo® Serum-Free Tumorsphere Media
Other Supplies Required
Reagents
Materials
- 15 mL conical tube
- Ultralow adhesion cell culture plates
Equipment
- 37 °C and 5% CO2 incubator
- Benchtop Centrifuge
Procedure Overview
Cryopreservation of Stem Cells
Add 2 U/mL Heparin and 0.5 µ;g/mL Hydrocortisone to pre-warmed StemXVivo® Serum-Free Tumorsphere Media.
Resuspend cells in the supplemented media.
Transfer the cell suspension to ultralow adhesion plates.
Incubate cultures in a 5% CO2 and 37 °C incubator.
Tumorspheres should form in 3-10 days.
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