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3-D Cell Culture Hydrogels

AS ONE INTERNATIONAL is proud to bring you two brands of distinct hydrogels for 3-D cell culture: Cellendes, a life science company in Germany, developed a biomimetic dextran-based hydrogel while Menicon Life Science in Japan manufactures a peptide-based hydrogel. Each hydrogel offers unique advantages for a variety applications like drug discovery and tissue engineering.

3-D Life Hydrogels from Cellendes

3-D Life hydrogel is a dextran-based hydrogel developed by Cellendes GmbH, a life science company offering a comprehensive technology for controlled design of cell environment in 3D cell culture. The 3-D Life Hydrogel backbone consists of dextran polymer functionalized with maleimides that are cross linked with thiol-bearing linear linkers, PEG-Link or CD-Link. The maleimide-dextran polymer and PEG-Link or CD-Link are mixed together at varying concentrations to adjust the elasticity of the gel. A soft gel may have an elastic shear modulus of 20 Pa while hard gels can reach 6,000 Pa. Hydrogels keep their elasticity at warmer and cooler temperatures and thus can be utilized in a wide range of applications.

3-D Life Hydrogels provide a neutral background matrix that is ideal for incorporating, suspending, and manipulating cells, tissues, biochemicals, and other entities in a “solid” aqueous-based environment or for creating a biologically inert elastic surface or coating. A toolbox of reagents provides flexibility in composing biomimetic hydrogels right at the researcher’s bench. Areas of application include basic research, drug development and biomedical engineering.

3-D Life Hydrogels are biocompatible allowing cells to be embedded during gel formation. Adhesion peptides can be incorporated into the hydrogel to adhere to your cell receptor. Dextranase added to cell culture medium or PBS will dissolve the hydrogel, releasing the cells. Cell integrity and physiology remain functional while in the gel and after gel dissolution.

Gel Characteristics

  • Adjustable elasticity– shear modulus from 20 – 6,000 Pa
  • High elasticity and low viscosity
  • Malleable – can be molded into any shape
  • Automation – aqueous solutions of hydrogel components allow microliter precision liquid handling
  • Rapid gelation keeps entities suspended in 3-D space
  • Scalability – from free-form gels to high throughput applications
  • Ambient temperature– gelation occurs at room temperature, no heating or cooling necessary
  • Translucent and biocompatible
  • Choice of biologically inert or biomimetically modified components

Structure

Maleimide-modified dextran polymer is the biologically inert basis for 3-D Life dextran hydrogels
Crosslinkers PEG or matrix metalloprotease cleavable CD-Link react with maleimide groups on the dextran polymer. Crosslinks polymers to form the gel.
Adhesion peptides (e.g. RGD Peptide) bind via their terminal cysteine to a portion of the maleimide groups of the dextran. Provides a cell adhesion matrix.
Cells bind to the dextran-peptide conjugates with their corresponding adhesion receptors (e.g. integrins)

Features and Benefits

Features Benefits
Defined composition of biologically inert synthetic polymers
  •  Nontoxic to cells
  • Does not activate or bind to cells
  • Dextranase dissolves the hydrogel allowing for the isolation of chemically fixed or living cells.
User-controlled modifications (e.g. peptides, proteins)
  • Add adhesion peptides – observe cellular response or create cell-type specific microenvironments
  • Add extracellular matrix proteins (e.g. fibronectin, laminin)
  • Add cleavable crosslinkers – observe migration and invasion
Robust gel formation and handling
  • Easy to use and no special equipment needed!
  • Adjust rate of gel formation
  • Embed single cells or cell spheroids
  • Cells remain suspended in the gel – no settling to the bottom.
Wide range of ligand density (up to 5 mmol/L)
  • Offers a high capacity for biomimetic modifications for the creation of complex microenvironments
Tunable gel stiffness
  • Adjust the gel elasticity to mimic your cells natural environment.

Choosing the right 3-D Life Hydrogel components

Potential Applications

  • Suspend biologics or other entities for manipulation
  • Suspend chemicals and biochemicals for slow releasing
  • Miniaturize your application with microarray hydrogels
  • Form an aqueous barrier between different materials or to block an area
  • Culture cells in or on top of the hydrogel
  • Create an elastic surface
  • Create a biocompatible coating to protect tissue sections
  • Create a protective coating to reduce sample dehydration
  • Automate or create your own cell migration assay
  • Drug discovery with 3D cell culture

Application Notes

Formation of Hydrogel after mixing of Mal-Dextran and PEG-Link at a final concentration of 3 mMol/L maleimide groups and 3 mMol/L thiol groups.

Soft gel Hard gel
Elastic shear modulus 20 -40 Pa 4,000 – 6,000 Pa
Conc. of thiol groups 1.5 mMol/L 6 mMol/L
Conc. of maleimide groups 1.5 mMol/L 6 mMol/L

3-D Life Hydrogel components Mal-Dextran and PEG-Link (Dextran-PEG), Mal-Dextran and CD-Link (Dextran-CD), Mal-PVA and PEG-Link (PVA-PEG) and Mal-PVA and CD-Link (PVA-CD), as indicated in the diagram, were mixed together at various final concentrations of reactive groups. The elastic shear modulus of each hydrogel was measured 20 min after the mixing of the polymer components. Each data point represents the average of three independent measurements.

Recent Publications

Ayenehdeh et al. Immunomodulatory and protective effects of adipose tissue-derived mesenchymal stem cells in an allograft islet composite transplantation for experimental autoimmune type 1 diabetes. Immunology Letters. 2017, 188: 21-31.

Martina et al. Modeling human immunity in vitro: improving artificial lymph node physiology by stromal cells. Applied In Vitro Toxicology. 2016, 2(3): 143-150.

Charwat et al. Potential and limitations of microscopy and Raman spectroscopy for live-cell analysis of 3D cell cultures. Journal of Biotechnology. 2015, 205: 70-81.

Order Now:

Cat # Name Size
09-G-001 3-D Life PVA-PEG Hydrogel Kit 1 kit*
09-P-001 3-D Life RGD Peptide 1 umol
09-P-003 3-D Life Scrambled RGD Peptide 1 umol
B10-3 3-D Life 10x CB Buffer (pH 5.5) 600 ul
B20-3 3-D Life 10x CB Buffer (pH 7.2) 600 ul
D10-1 3-D Life Dextranase 500 ul
G81-1 3-D Life PVA-CD Hydrogel Kit 1 kit
G82-1 3-D Life PVA-PEG Hydrogel SG (Slow Gelling) 1 kit
G83-1 3-D Life PVA-CD Hydrogel SG (Slow Gelling) 1 kit
G90-1 3-D Life Dextran – PEG Hydrogel Kit 1 kit
G91-1 3-D Life Dextran – CD Hydrogel Kit 1 kit
G92-1 3-D Life Dextran-PEG Hydrogel SG (Slow Gelling) 1 kit
G93-1 3-D Life Dextran – CD Hydrogel SG (Slow Gelling) 1 kit
G94-1 3-D Life ToGro 1 kit
L50-1 3-D Life PEG – Link 200 ul
L50-3 3-D Life PEG – Link 3 x 200 ul
L60-1 3-D Life CD – Link 200 ul
L60-3 3-D Life CD – Link 3 x 200 ul
M80-3 3-D Life Maleimide-PVA Set 1 kit
M90-3 3-D Life Maleimide – Dextran 1 set
P10-3 3-D Life RGD Peptide 3 x 1 umol
P11-3 3-D Life Scrambled RGD Peptide 3 x 1 umol
T10-1 3-D Life Thioglycerol 60 ul

*Includes maleimide-dextran, crosslinker, buffers, dextranase, thio, water

PanaceaGel from Menicon Life Science

Menicon developed PanaceaGel as a synthetic alternative for collagen scaffolding commonly used in tissue engineering. PanaceaGel is a 13 amino acid peptide that self assembles to form beta sheets. The beta sheets stack on top of each other to form nanofibers. It is the network of interacting nanofibers that form PanaceaGel.

Structure and Mechanism

The main component of PanaceaGel is a thirteen amino acid synthetic peptide, which is made of four naturally occurring amino acids in a specific sequence (US 2012/0058066 A1). The primary structure is:

The thirteen amino acid peptides undergo spontaneous self-assembly in several steps to form nanofibers in aqueous solvents due to hydrogen bonding, electrostatic interaction and hydrophobic interaction. These nanofibers assemble into a higher-order structure, a nanofiber-scale three-dimensional network, which forms a transparent gel with a moisture content of over 99%.

 

Gel formation by self-assembly

Step 1: Beta sheet structure – 13 amino acid peptides form beta sheets by hydrogen bonding and electrostatic and hydrophobic interactions.

Step 2: Nanofiber formation – Each beta sheet has 2 sides, a hydrophilic surface and a hydrophobic surface. Beta sheets stack on top of each other to form nanofibers in an aqueous solvent (water) by hydrophobic interactions.

Step 3: Nanofiber network structure – Nanofibers interact with each other to form a network containing more than 99% water.

Features and Benefits

Features Benefits
Self assembly 13 amino acid peptides
  • Ready to use right out of the vial!
  • No buffers required
  • Dilute hydrogel in culture medium, add cells
100% synthetic polymer
  • Synthetic substitution for collagen scaffolding
  • No animal derived components
  • No risk of pathogen contamination
Lot to lot consistency
  • Reproducible data
Neutral pH
  • Easy on cells
  • Ideal for 3-D scaffolding
  • Carrier for drug delivery
Stable at room temperature
  • No special storage conditions
  • Long shelf life
  • Easy to work with

Recent Publications

Nakamichi et al. Mohawk promotes the maintenance and regeneration of the outer annulus fibrosus of intervertebral discs. Nature Communications, 2016, 7: 12503.

Nagai et al. The mechanical stimulation of cells in 3D culture within a self-assembling peptide hydrogel. Biomaterials, 2012, 33(4): 1044-1051.

Uehara et al. Exogenous fatty acid binding protein 4 promotes human prostate cancer cell progression. Int. J. Cancer, 2014, 135: 2558–2568.

Order Now:

Cat.# Component Size
SPG-178-004 PanaceaGel, 0.4%  1 ml
SPG-178-104 PanaceaGel, 0.4%, isotonic  1 ml
SPG-178-008 PanaceaGel, 0.8%  1 ml
SPG-178-108 PanaceaGel, 0.8%, isotonic  1 ml