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PNIPAAm-PEG a block co-polymer of poly(N-isopropylacrylamide) and poly(ethylene glycol) for 3D Cell Culture and other applications

Mebiol® Gel PNIPAAm-PEG 3D Thermoreversible Hydrogel

Background


Mebiol® Gel 
[2477 kb]
 

Hydrogels are a diverse class of polymeric materials characterized by their network-like structure and high water content. Hydrogels of many kinds have found a wide variety of applications in medicine and life science research weighted towards, but not at all limited to three-dimensional cell culture, tissue engineering, and drug delivery. Properties highly favorable to cell culture and tissue engineering applications prompted the commercialization of Mebiol® Gel, a copolymer of poly(N-isopropylacrylamide) and poly(ethylene glycol) (PNIPAAm-PEG) for research purposes in the early 2000's.

Mebiol® Gel's defining feature, in contrast to other commercially available hydrogels, is its temperature reversible sol-gel transition. When cooled, Mebiol® Gel is a sol (handles like a liquid) but becomes a rigid hydrogel at higher temperatures. In practice, this means extremely easy cell handling. Cultures are seeded into cooled Mebiol® Gel and recovered conveniently by cooling the culture vessel and centrifugation. In the gel state, the highly lipophylic environment of the Mebiol® Gel presents an efficient niche for cell proliferation, cell communication, gas and mass exchange, and protection of cells and tissue from shear forces.

Low Temperature (Sol) High Temperature (Gel)

Features

· Easy handling

· Non-toxic, biocompatible

· 100% synthetic, pathogen free

· High transparency for cell observation

· Proven performance.

Preparation

Mebiol Gel Preparation

1. Add Culture Medium

2. Dissolve Mebiol® Gel

3. Cultivation of Cells / Tissues

4. Recovery of Cells / Tissues

Application

· Stem cell and pluripotent stem cell culture, expansion, and differentiation

· Spheroid culture

· Cell implantation

· Organ and Tissue Regeneration

· Drug Delivery

· Non-cell culture application

Freeze-drying Polymer is put in each flask as follows. 

  • MBG-PMW20-1001 (10 mL) : 1g
  • MBG-PMW20-5001 (50 mL) : 5g

Application Examples

1) Culture of primary cancer cells in Mebiol Gel
Selective growth of only primary cancer cells from human cancerous tissue in Mebiol Gel (courtesy Dr. S. Kubota, Dept. of General Surgery, St. Marianna University School of Medicine). This technology enables the characterization of patient-derived primary cancer cells and therefore enabling the evaluation of primary cells for chemosensitivity, malignancy, metastasis activity and other parameters that might influence patient therapy.

Human colon cancerous tissue was cultured in Mebiol® Gel for 10 days. Only primary cancer cells proliferate from the tissue in Mebiol Gel.

Fibroblasts growth in Mebiol® Gel is suppresed whereas In collagen and many other 3D gel culture matrices, fibroblasts overgrow and prevent proliferation of primary cancer cells.

2) Stem Cell Culture
3D culture of undifferentiated mouse and Macaca ES cells cultured without LIF or feeder layer cells performed in collaboration with with Dr. K. Hishikawa, Dept. of Clinical Renal Regeneration, University of Tokyo. 

Left: 2D on Feeder Cells
Right: 3D Culture in Mebiol Gel
(Day 7)

The strong positive alkaline phosphatase staining of Macaca (primate) ES cells cultured in Mebiol® Gel suggests undifferentiation.

Left: 2D on Feeder Cells
Right: 3D Culture in Mebiol Gel
(without LF, day 5)

 
3) Selective separate culture of somatic stem cell (mouse embryo skin origin)
Isolation of Epithelial Stem Cells from Dermis by a Three-dimensional Culture System
Journal of Cellular Biochemistry, 98 (1), 174-184 (2006) PMID: 16408300
 
4) Cartilage tissue reproduction by in vitro 3D culture of cartilage cell
Chondrocytes Containing Growth Factors in a Novel Thermoreversible Gelation Polymer Scaffold
Tissue Engineering, 12 (5), 1237-1245 (2006)
 
5) Bone induction by in vitro 3D culture of human mesenchymal cell stem cell (hMSC)
Gene expression profile of human mesenchymal stem cells during osteogenesis in three-dimensional thermoreversible gelation polymer
Biochem. Biophys. Res. Commun., 317, 1103-1107 (2004). PMID: 15094382
 
6) Production of hepatitis C viruses (HCV) by 3-D culture of human hepatocyte cell line
Production of infectious hepatitis C virus particles in three-dimensional cultures of the cell line carrying the genome-length dicistronic viral RNA of genotype 1b
Virology, 351 (2), 381-392 (2006) PMID: 16678876
 
7) Passage control by local heating (on chip cell sorter system)
On-Chip Cell Sorting System Using Laser-Induced Heating of a Thermoreversible Gelation Polymer to Control Flow, Y. Shirasaki, J. Tanaka, H. Makazu, K. Tashiro, S. Shoji, S. Tsukita, T. Funatsu,
Anal. Chem., 78, 695-701 (2006) PMID: 16448041
more

Technical Notes

 Mebiol® Gel Literature Citations and References

 Mebiol® Gel Frequently Asked Questions (FAQ)

Instruction Manual for Mebiol Gel PDF

Dynamic Viscoelastic Property of Mebiol® Gel


Temperature dependence of the dynamic moduli of the aqueous solution of Mebiol® Gel at a concentration of 10 wt% in distilled water. Storage modulus (G', solid lines) and loss modulus (G'', broken lines) were measured on heating (closed symbols) and cooling (open symbols) at the oscillatory frequency of 1 Hz.

Product List

Product Name Cat# Quantity Price

Mebiol Gel

MBG-PMW20-1001 1*10ML

¥ 30,000
$ 400
€ 300

Mebiol Gel

MBG-PMW20-5001 1*50ML

¥ 60,000
$ 800
€ 600

Mebiol Gel

MBG-PMW20-1005 5*10ML

¥ 100,000
$ 1334
€ 1000

Mebiol Gel

MBG-PMW20-5005 5*50ML

¥ 200,000
$ 2667
€ 2000

To be used for research only. DO NOT use for human gene therapy or clinical diagnosis.

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