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Nissan Chemical Industries, Ltd.
New three-dimensional culture medium FCeM® series

Overview

FCeM® is a three-dimensional culture medium containing a combination of polymer "FP001" which uniformly disperses the cells clumps.
By the effect of the FP001-containing medium,  it maintains / promotes the growth and function of cells in a non-adhesive state.

It enables the suspension culture of spheres (spheroids).
It suppresses excessive agglomeration of spheres.
 It improves proliferative capacity and gets  the original function of cells
 It improves penetration of drugs and nutritional ingredients
It allows to create a sphere from a single cell
Viscosity is equivalent to water, easy to add compound
Animal-derived materials such as BSA are not included

1. Features

FCeM® uniformly disperses cell clumps while showing the same viscosity as water.
It makes it possible to culture floating spheres (cell mass) at a low concentration (0.015%).

【The growth of cancer cells (cultured for 5 days)】
FCeM® suppresses the formation of giant clumps by A549 and HCT116 and forms a uniform spheres.

 

【The nature of the medium containing FP001】
Easy to handle  as viscosity is equivalent to water and the conventional medium

  Additive-free FP001
0.015%
Methylcellulose
0.6%
Collagen
0.2%
Viscosity [mPa・s]  1 1 12
Sedimentation [Pa] 0.02 7 10
HepG2 Precipitating Floating Floating Floating
Sphere   Dispersion Dispersion Dispersion

 

【Actually used cells 】
FCeM® series can be used to a variety of cancer cells.

Cell name Origin FP001
A549 Lung Favorable
A375 Melanoma Favorable
HCT116 Colon Favorable
HeLa Uterus Favorable
MCF7 Breast Favorable
MDA-M B-231 Breast Favorable
Cell name Origin FP001
HepG2 Liver Favorable
MNNG/NOS Osteosarcoma Favorable
AGS Stomach Favorable
MIAPaCa-2 Pancreas Favorable
A431 Skin Favorable *
SKOV3 Ovary Favorable *
                                                                            * HB-EGF addition

2. Confirmation of anticancer drug sensitivity

【Effects of existing anti-cancer drugs】
Comfirmation of  s sensitivity of various anti-cancer drugs by using the FP001-containing medium

【FP001 / 384well low adhesive plate three-dimensional culture】
We also confirmed the sensitivity of anti-cancer drugs in 384well plate, and made sure that this is a test system which can withstand even in HTS assessment.

3. Non-adherent culture of cancer cells

Concerning proliferation of A549 cells and HCT116 cells on low attachment plates Using FCeMR, the number of cells (WST value) in FCeMTM was 10 times higher at the maximum compared to that in the control culture medium.

4. Effects of HB-EGF to human ovarian cancer · SKOV3 cell proliferation

We confirmed the growth of HB-EGF dependence in SKOV3 cells ⇒ which enables to optimize the evaluation system by the addition of growth factors

5. Anchorage-independent growth

【Evaluation of specific anchorage-independent growth by growth factors】
In culture method which combines low attachment plate and FP001, the anchorage-independent growth of human ovarian cancer-derived SKOV3 cells by EGF receptor ligand and TGF-β can be evaluated.

 

【Inhibitory effect of Gefitinib against the anchorage-independent growth by HB-EGF
In culture method which combines low attachment plate and FP001,  against the anchorage-independent growth by HB-EGF,Gefitinib as an EGF receptor inhibitor exhibits specific inhibitory effects.

【Susceptibility test for Hela, A549 of Trametinib】
In the three-dimensional culture using FCeM®-D medium, the sensitivity of the EGF signaling pathway inhibitor increases.

Cell IC50 (nM)
Two-dimensional culture FP001 Three-dimensional culture
HeLa > 30 2.4
A549 18.2 3.4

6. Product List

Product Name Base medium Package Size Wako catalog No. Nissan Chemical's product No. Storage Condition
            FCeM®-D D-MEM 500 mL 387-06265 SJ000546 Keep at 2-10°C.
            FCeM®-R RPMI-1640 500 mL 380-06255 SJ000545
            FCeM®-D/F D-MEM/Ham'sF12 500 mL 383-06245 SJ000500

7. Related Products

        They are kits to prepare FCeM® medium with your medium in stock. You can prepare the 50 mL medium using the kit.
Product Name Package Size Wako catalog No. Storage Condition
            FCeM-series Preparation kit 1 test 385-07981 Keep at 2-10°C.
            FCeM-series Preparation kit for Stem Cells 1 tests 382-07991
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