What is the composition of the medium used to culture MCF-7 cells for the data shown on the StemXVivo Media supplement (100X), Catalog # CCM017 product insert?

The medium used to cuture MCF-7 cells for the data shown on Catalog # CCM017 is MEM containing NEAA, Sodium Pyruvate, 10% FBS, 2mM L-Glutamine, and Penicillin-Streptomycin

Has StemXVivo Media supplement (100X), Catalog # CCM017, been tested on mouse epithelial cells?

Elaborate testing of Catlaog # CCM017 with mouse epithelial cells has not been performed. The supplement was tested once with NMuMG mouse mammary epithelial cells and found to induce EMT.

StemXVivo EMT Inducing Media Supplement (100X)

Name: StemXVivo EMT Inducing Media Supplement (100X)
Brand: R&D Systems
SKU: CCM017
Rating: 5 (6 reviews)

R&D Systems, Inc. a Bio-Techne Brand

Catalog # CCM017

Citations (8)

(6)

Product Datasheet / COA / SDS

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Setting the standard in quality research reagents

Summary for StemXVivo EMT Inducing Media Supplement (100X)

Kit Summary

To reliably induce epithelial to mesenchymal transition (EMT) in vitro.

Key Benefits

Induces EMT in only 5 days
Is compatible with multiple cell types
Is a defined formulation to provide consistent EMT induction

Why Induce EMT in vitro with a Defined Media Supplement?

Designed to study EMT in vitro, this media supplement contains a cocktail of EMT-inducing recombinant proteins and neutralizing antibodies for the straightforward induction of EMT in a variety of cell types.

Reliable induction of EMT is useful for understanding the biology of EMT and for identifying and evaluating novel prognostic markers and therapeutics for fibrotic diseases and metastasis. However, the ability to induce EMT in vitro typically requires TGF-beta stimulation or labor-intensive genetic modification. Moreover, although TGF-beta is necessary to induce EMT, it is not sufficient in all cell types.

StemXVivo^® EMT Inducing Media Supplement:

Drives EMT in only 5 days to ensure efficient use of time and reagents.
Provides a straightforward method to induce EMT in vitro.
Is completely defined to reduce experimental variability.
May induce EMT in cells resistant to TGF-beta stimulation alone.
Avoids time-consuming genetic modifications.

Kit Contents

Components

This defined media supplement contains the following premium quality proteins and high performance neutralizing antibodies to induce EMT:

Recombinant Human Wnt-5a
Recombinant Human TGF-beta1
Anti-Human E-Cadherin
Anti-Human sFRP-1
Anti-Human Dkk-1

Each vial contains sufficient volume to drive EMT in the equivalent of fifteen 10 cm diameter tissue culture plates and has been shown to induce EMT in the following human cell lines:

MCF-7 human breast cancer cells
MCF-10A human breast epithelial cells
HT-29 human colon adenocarcinoma cells
A549 human lung carcinoma cells
A431 human epithelial carcinoma cells

Epithelial to Mesenchymal Transition (EMT) is a biological process which is centrally important to embryogenesis and organ development. Epithelial are highly ordered monolayers of cells with a uniform morphology. Cells within epithelial are characterized by the fact that they are adhered tightly to each other. In contrast, mesenchymal cells differ in shape and display an increased capacity for migration and invasion. This change in phenotype is thought to be involved in some oncogenic pathways. Epithelial to mesenchymal transition allows benign tumors to progress into metastatic cancers that can invade other tissues. EMT is also involved in fibrosis during scar tissue development and may be pathologically relevant to the development of progressive fibrotic diseases. Molecular markers of epithelial to mesenchymal transition include increased expression of N-Cadherin and Vimentin, nuclear localization of beta-catenin, and augmented levels of transcription factors that reduce E-Cadherin expression (i.e. Snail1).

Product Specifications for StemXVivo EMT Inducing Media Supplement (100X)

Species

Human

Scientific Data Examples for StemXVivo EMT Inducing Media Supplement (100X)

	Induction of Epithelial to Mesenchymal Transition(EMT) in the A549 human lung carcinoma cell line. The A549 human lung carcinoma cell line was cultured without(A, B); or with(C, D)the StemXVivo^®EMT Inducing Media Supplement (Catalog #CCM017) for five days.(A)Control cells exhibit typical epithelial morphology compared to the spindle-shaped, mesenchymal morphology observed in cells following EMT induction(C). EMT status was also assessed by dual immunofluorescence.(B)Untreated cells express the epithelial marker E-Cadherin (red) and exhibit little labeling of the mesenchymal marker Fibronectin (green).D)In contrast, EMT induction results in decreased E-Cadherin expression (red) and an increase in Fibronectin labeling (green). E-Cadherin was detected in cells using a NorthernLights™ (NL) 577-conjugated Goat Anti-Human E-Cadherin Antigen Affinity-purified Polyclonal Antibody (Catalog # NL648R; red). Fibronectin was detected using a Mouse Anti-Human Fibronectin Monoclonal Antibody (Catalog # MAB1918), followed by a NL493-conjugated Donkey Anti-Mouse IgG Secondary Antibody (Catalog # NL009; green). The nuclei were counterstained with DAPI (blue).
	Upregulation of the Mesenchymal Markers, Snail and Vimentin, in EMT-induced Cells. A549 human lung carcinoma and MCF 10A human breast epithelial cells were either untreated or cultured with the StemXVivo^®EMT Inducing Media Supplement (Catalog # CCM017) for five days. The cells were analyzed for an epithelial to mesenchymal transition (EMT) by simultaneously staining with the antibodies contained in the Human EMT 3-Color Immunocytochemistry Kit (Catalog # SC026): NorthernLights™ (NL) 637-conjugated Goat Anti-Human E-Cadherin (pseudocolored white), NL557-conjugated Goat Anti-Human Snail (red), and NL493-conjugated Rat Anti-Human Vimentin (green). Cells cultured with the EMT Inducing Media Supplement showed downregulation of the epithelial marker, E-Cadherin, and concurrent upregulation of the mesenchymal markers, Snail and Vimentin, compared to control cells.

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, an epithelial to mesenchymal transition (EMT) is induced using the following in vitro differentiation procedure:

Add EMT Inducing Media Supplement directly to the standard culture media of your cells of interest
Culture cells for only 5 days to induce EMT
Evaluate EMT status using characteristic morphological changes and marker antibodies

View Graphic Procedure

Reagents Provided

Reagents supplied in the StemXVivo^® EMT Inducing Media Supplement (Catalog # CCM017):

1 mL of StemXVivo^® EMT Inducing Media Supplement

Note: The quantity of media supplement is sufficient to drive EMT in the equivalent of fifteen 10 cm diameter tissue culture plates.

Other Supplies Required

Reagents

Cell culture medium
Dissociation solution (e.g., TrypLE™ Express, Invitrogen^® or equivalent)
0.4% Trypan blue solution

Materials

Epithelial cells of interest
Tissue culture plates/flasks
15 mL centrifuge tubes
Pipettes and pipette tips
Serological pipettes

Equipment

37 °C and 5% CO₂ incubator
Centrifuge
Hemocytometer
Inverted microscope
37 °C water bath
2 °C to 8 °C refrigerator

Procedure Overview

Harvest epithelial cells of interest using a dissociation solution.

<strong>Harvest</strong> epithelial cells of interest using a dissociation solution

Transfer the cell suspension to a 15 mL conical tube containing pre-warmed media.

Centrifuge at 400 x g for 5 minutes.

Perform a cell count.

Plate cells at 0.9 - 1.0 x 10⁴ cells/cm² in media containing 1X EMT Inducing Media Supplement.

Plate cells at 0.9 - 1.0 x 10 4 cells/cm2 in media containing 1X EMT Inducing Media Supplement.

Culture cells and monitor cell morphology daily.

Replace media after 3 days.

After 5 days, EMT induction is complete.

Evaluate EMT status using established markers and immunocytochemistry

Citations for StemXVivo EMT Inducing Media Supplement (100X) (8)

Showing 1 - 8 of 8 citations Showing All

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Citations are publications that use Bio-Techne products. Selected citations for StemXVivo EMT Inducing Media Supplement (100X) include:

Species: Human
Sample Types: Whole Cells
Applications: Cell Culture

B Ghosh et al. (2022), Epithelial plasticity in COPD results in cellular unjamming due to an increase in polymerized actin Journal of Cell Science, 0(0).
PMID: 35118497
Species: Human
Sample Types: Whole Cells

C Kötting et al. (2021), Immune-Stimulatory Effects of Curcumin on the Tumor Microenvironment in Head and Neck Squamous Cell Carcinoma Cancers, 13(6).
PMID: 33809574
Species: Human
Sample Types: Whole Cells
Applications: Cell Culture

HX Dang et al. (2020), Long non-coding RNA LCAL62 / LINC00261 is associated with lung adenocarcinoma prognosis Heliyon, 6(3):e03521.
PMID: 32181394
Species: Human
Sample Types: Whole Cells

YL Liu et al. (2019), Assessing metastatic potential of breast cancer cells based on EGFR dynamics Sci Rep, 9(1):3395.
PMID: 30833579
Species: Human
Sample Types: Whole Cells
Applications: Bioassay

KK Brodaczews et al. (2019), Metastatic renal cell carcinoma cells growing in 3D on poly?D?lysine or laminin present a stem?like phenotype and drug resistance Oncol. Rep., 42(5):1878-1892.
PMID: 31545459
Species: Human
Sample Types: Whole Cells
Applications: Bioassay

I Druzhkova et al. (2018), E-Cadherin in Colorectal Cancer: Relation to Chemosensitivity Clin Colorectal Cancer, 0(0).
PMID: 30415989
Species: Human
Sample Types: Whole Cells
Applications: Bioassay

A Siddiqui et al. (2017), Thymidylate synthase is functionally associated with ZEB1 and contributes to the epithelial-to-mesenchymal transition of cancer cells J. Pathol, 0(0).
PMID: 28337746
Species: Human
Sample Types: Whole Cells

Scheel C et al. (2011), Paracrine and autocrine signals induce and maintain mesenchymal and stem cell states in the breast. Cell, 145(6):926-40.
PMID: 21663795

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Customer Reviews for StemXVivo EMT Inducing Media Supplement (100X) (6)

4.5 out of 5

6 customer ratings

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StemXVivo EMT Inducing Media Supplement (100X)

Name: Anonymous

Verified Customer | Posted 08/31/2021
StemXVivo EMT Inducing Media Supplement (100X)

Name: Anonymous

Verified Customer | Posted 02/20/2021
StemXVivo EMT Inducing Media Supplement (100X)

Name: Anonymous

Verified Customer | Posted 04/08/2019
StemXVivo EMT Inducing Media Supplement (100X)

Name: Leslie Priddy

Verified Customer | Posted 04/04/2018
StemXVivo EMT Inducing Media Supplement (100X)

Name: Babak Nami

Verified Customer | Posted 08/08/2017

It seems very useful, but too expensive. I gave up using it because of the high cost.
StemXVivo EMT Inducing Media Supplement (100X)

Name: Anonymous

Verified Customer | Posted 04/12/2017

StemXVivo EMT Inducing Media Supplement was used to induce EMT of ARPE-19 cells. Morphology and fibronectin measured by fibronectin ELISA (DY1918-05 and DY008, R & D Systems) are attached.

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FAQs for StemXVivo EMT Inducing Media Supplement (100X)

What is the composition of the medium used to culture MCF-7 cells for the data shown on the StemXVivo Media supplement (100X), Catalog # CCM017 product insert?

The medium used to cuture MCF-7 cells for the data shown on Catalog # CCM017 is MEM containing NEAA, Sodium Pyruvate, 10% FBS, 2mM L-Glutamine, and Penicillin-Streptomycin
Has StemXVivo Media supplement (100X), Catalog # CCM017, been tested on mouse epithelial cells?

Elaborate testing of Catlaog # CCM017 with mouse epithelial cells has not been performed. The supplement was tested once with NMuMG mouse mammary epithelial cells and found to induce EMT.

StemXVivo EMT Inducing Media Supplement (100X)