StemXVivo Serum-Free Tumorsphere Media

For Tumorsphere Formation of Human/Mouse/Rat Cancer Cells
Catalog # Availability Size / Price Qty
Product Details
Citations (8)
Supplemental Products

StemXVivo Serum-Free Tumorsphere Media Summary

Kit Summary

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.


This product contains human transferrin. The transferrin was 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. As no testing can offer complete assurance of freedom from infectious agents, this product should be handled as if capable of transmitting infection.


  • 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.


Data Examples
Tumorspheres Formed from the MCF-7 Human Breast Cancer Cell Line
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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 104 cells per well in a 6-well ultralow adhesion culture plate and cultured at 37 °C and 5% CO2 for 3-10 days to induce tumorsphere formation. Tumorspheres were imaged using a CloneSelect Imager from Molecular Devices (New Milton) Limited.

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.


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

Product Datasheets

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


  • Heparin (Tocris Catalog # 2812)
  • Hydrocortisone (Tocris Catalog # 4093)
  • Cell dissociation solution


  • 15 mL conical tube
  • Ultralow adhesion cell culture plates


  • 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.

Add 2 U/mL Heparin and 0.5 ug/mL Hydrocortisone to pre-warmed StemXVivo Serum-Free Tumorsphere Media.

Resuspend cells in the supplemented media.

Resuspend cells in the supplemented media.

Transfer the cell suspension to ultralow adhesion plates.

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.

Tumorspheres should form in 3-10 days.

Reagents Provided

Other Reagents Provided

Citations for StemXVivo Serum-Free Tumorsphere Media

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

8 Citations: Showing 1 - 8
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  1. RER1 enhances carcinogenesis and stemness of pancreatic cancer under hypoxic environment
    Authors: S Chen, J Zhang, J Chen, Y Wang, S Zhou, L Huang, Y Bai, C Peng, B Shen, H Chen, Y Tian
    J. Exp. Clin. Cancer Res., 2019;38(1):15.  2019
  2. Metastatic renal cell carcinoma cells growing in 3D on poly?D?lysine or laminin present a stem?like phenotype and drug resistance
    Authors: KK Brodaczews, ZF Bielecka, K Maliszewsk, C Szczylik, C Porta, E Bartnik, AM Czarnecka
    Oncol. Rep., 2019;42(5):1878-1892.  2019
  3. Depletion of HDAC1, 7 and 8 by Histone Deacetylase Inhibition Confers Elimination of Pancreatic Cancer Stem Cells in Combination with Gemcitabine
    Authors: MH Cai, XG Xu, SL Yan, Z Sun, Y Ying, BK Wang, YX Tu
    Sci Rep, 2018;8(1):1621.  2018
  4. The Antihelminthic Niclosamide Inhibits Cancer Stemness, Extracellular Matrix Remodeling, and Metastasis through Dysregulation of the Nuclear ?-catenin/c-Myc axis in OSCC
    Authors: LH Wang, M Xu, LQ Fu, XY Chen, F Yang
    Sci Rep, 2018;8(1):12776.  2018
  5. Role of SIRT2 in Regulation of Stemness of Cancer Stem-Like Cells in Renal Cell Carcinoma
    Authors: R Wei, D He, X Zhang
    Cell. Physiol. Biochem., 2018;49(6):2348-2357.  2018
  6. TRPM7 overexpression enhances the cancer stem cell-like and metastatic phenotypes of lung cancer through modulation of the Hsp90?/uPA/MMP2 signaling pathway
    Authors: K Liu, SH Xu, Z Chen, QX Zeng, ZJ Li, ZM Chen
    BMC Cancer, 2018;18(1):1167.  2018
  7. Oncostatin M induces tumorigenic properties in non-transformed human prostate epithelial cells, in part through activation of signal transducer and activator of transcription 3 (STAT3)
    Authors: S Sterbova, T Karlsson, E Persson
    Biochem. Biophys. Res. Commun., 2018;0(0):.  2018
  8. HCMV activates the IL-6-JAK-STAT3 axis in HepG2 cells and primary human hepatocytes.
    Authors: Lepiller Q, Abbas W, Kumar A, Tripathy M, Herbein G
    PLoS ONE, 2013;8(3):e59591.  2013


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