Can a 96-well ultra-low adhesion plate be used for Tumorsphere formation using Catalog # CCM012 (StemXVivo Serum-free Tumorsphere Media)? How many cells should be plated /well?

Name: StemXVivo Serum-Free Tumorsphere Media
Brand: R&D Systems
SKU: CCM012

Yes. A 96-well ultra-low adhesion plate can be used for Tumorsphere formation. A recommended cell number is approximately 100 cells/well, but it is advisable to titrate and evaluate the optimal cell number.

StemXVivo Serum-Free Tumorsphere Media

Catalog # CCM012 | R&D Systems, Inc. a Bio-Techne Brand

Citations (13)

Product Datasheet / COA / SDS

For Tumorsphere Formation of Human/Mouse/Rat Cancer Cells

Key Product Details

Setting the standard in quality research reagents

Summary for StemXVivo Serum-Free Tumorsphere Media

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.

Precautions

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.

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.

Product Specifications for StemXVivo Serum-Free Tumorsphere Media

Species

Human

Scientific Data Examples for StemXVivo Serum-Free Tumorsphere Media

	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.

Preparation & Storage

Shipping Conditions	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.

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

View Graphic Procedure

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

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

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.

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.

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.

Citations for StemXVivo Serum-Free Tumorsphere Media (13)

Showing 1 - 10 of 13 citations Showing All

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Citations are publications that use Bio-Techne products. Selected citations for StemXVivo Serum-Free Tumorsphere Media include:

Species: Mouse
Sample Types: Whole Cells

P Gonzalez-C et al. (2023-02-03), Cancer stem cell-derived extracellular vesicles preferentially target MHC-II-macrophages and PD1+ T cells in the tumor microenvironment PLoS ONE, 2023-018(2):e0279400.
PMID: 36735677
Species: Human
Sample Types: Whole Cells
Applications: Bioassay

Z Nehme et al. (2022-05-18), Polyploid giant cancer cells, EZH2 and Myc upregulation in mammary epithelial cells infected with high-risk human cytomegalovirus EBioMedicine, 2022-080(0):104056.
PMID: 35596973
Species: Human
Sample Types: Organoid
Applications: Bioassay

MJ Munro et al. (2021-08-24), Colon adenocarcinoma-derived cells possessing stem cell function can be modulated using renin-angiotensin system inhibitors PLoS ONE, 2021-016(8):e0256280.
PMID: 34428252
MJ Munro et al. (2020-05-19), Colon adenocarcinoma-derived cells that express induced-pluripotent stem cell markers possess stem cell function PLoS ONE, 2020-015(5):e0232934.
PMID: 32428045
Species: Human
Sample Types: Whole Cells
Applications: Bioassay

V Yoganandar et al. (2020-01-30), Identification of Cancer Stem Cell Subpopulations in Head and Neck Metastatic Malignant Melanoma Cells, 2020-09(2).
PMID: 32019273
Species: Human
Sample Types: Whole Cells
Applications: Bioassay

KK Brodaczews et al. (2019-09-18), Metastatic renal cell carcinoma cells growing in 3D on poly?D?lysine or laminin present a stem?like phenotype and drug resistance Oncol. Rep., 2019-042(5):1878-1892.
PMID: 31545459
Species: Human
Sample Types: Whole Cells

S Chen et al. (2019-01-10), RER1 enhances carcinogenesis and stemness of pancreatic cancer under hypoxic environment J. Exp. Clin. Cancer Res., 2019-038(1):15.
PMID: 30630537
Species: Human
Sample Types: Whole Cells

K Liu et al. (2018-11-26), TRPM7 overexpression enhances the cancer stem cell-like and metastatic phenotypes of lung cancer through modulation of the Hsp90?/uPA/MMP2 signaling pathway BMC Cancer, 2018-118(1):1167.
PMID: 30477473
Species: Mouse
Sample Types: Transfected Whole Cells
Applications: Flow Cytometry

R Wei et al. (2018-09-27), Role of SIRT2 in Regulation of Stemness of Cancer Stem-Like Cells in Renal Cell Carcinoma Cell. Physiol. Biochem., 2018-049(6):2348-2357.
PMID: 30261494
Species: Human
Sample Types: Whole Cells

LH Wang et al. (2018-08-24), The Antihelminthic Niclosamide Inhibits Cancer Stemness, Extracellular Matrix Remodeling, and Metastasis through Dysregulation of the Nuclear ?-catenin/c-Myc axis in OSCC Sci Rep, 2018-08(1):12776.
PMID: 30143678
Species: Human
Sample Types: RWPE-1

S Sterbova et al. (2018-03-12), 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) Biochem. Biophys. Res. Commun., 2018-00(0).
PMID: 29526757
Species: Human
Sample Types: Whole Cells

MH Cai et al. (2018-01-26), Depletion of HDAC1, 7 and 8 by Histone Deacetylase Inhibition Confers Elimination of Pancreatic Cancer Stem Cells in Combination with Gemcitabine Sci Rep, 2018-08(1):1621.
PMID: 29374219
Species: Human
Sample Types: Whole Cells

Lepiller Q et al. (2013-03-26), HCMV activates the IL-6-JAK-STAT3 axis in HepG2 cells and primary human hepatocytes. PLoS ONE, 2013-08(3):e59591.
PMID: 23555719

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FAQs for StemXVivo Serum-Free Tumorsphere Media

Can a 96-well ultra-low adhesion plate be used for Tumorsphere formation using Catalog # CCM012 (StemXVivo Serum-free Tumorsphere Media)? How many cells should be plated /well?

Yes. A 96-well ultra-low adhesion plate can be used for Tumorsphere formation. A recommended cell number is approximately 100 cells/well, but it is advisable to titrate and evaluate the optimal cell number.