Easily Recover Live or Fixed Cells from Hydrogels

Do you need a hydrogel for 3D cell culture that allows for protease-free, post-culture recovery of live or chemically fixed cells?

3-D Life Dextranase

3-D Life dextran-based hydrogels can be dissolved by addition of 3-D Life Dextranase. This allows the isolation of chemically fixed or live cells after 3D cell culture for downstream processing of cells.

For example, cells can be passaged for further growth or analyzed by immunostaining, RT-PCR, biochemical analyses or flow cytometry, similar to cells from 2D cell cultures.

Since dextranase specifically degrades the dextran polymer meshwork without protease activity, cells and extracellular matrix are left intact during gel dissolution.

3-D Life Hydrogel Technology

The 3-D Life technology is an integrated system for the generation of biomimetic hydrogels used in 3D cell culture. The 3-D Life system consists of two polymers that form a hydrogel by covalent crosslinking at ambient temperature. The chemical reaction is non-toxic to cells and occurs rapidly, preventing cell settlement before the gel is formed.

Setup of 3-D Life biomimetic hydrogels for 3D cell culture

Features

  • Defined composition of biologically inert synthetic polymers and biopolymers
  • User-controlled biomimetic modifications (e.g. peptides, proteins)
  • Robust gel formation and handling
  • Wide range of ligand density (up to 5 mmol/l)
  • Tunable gel stiffness
  • Protease-free dissolution of gels for easy cell recovery

Dextran-based hydrogels

  • ToGro Hydrogel Kit: a Ready-To-Go kit that contains reagents for the preparation of hydrogels with a soft stiffness (400 Pa; shear modulus) and a RGD Peptide concentration of 0.4 mmol/L. The gel composition allows three-dimensional spreading and migration of most cells.
  • Dextran-PEG Hydrogel FG Kit: a Ready-To-Design kit that provides reagents for setting up fast gelling, cell-compatible hydrogels. Its major components are maleimide-functionalized dextran (Mal-Dextran) and the crosslinker PEG-Link.
  • Dextran-CD Hydrogel FG Kit: a Ready-To-Design kit that provides reagents for setting up fast gelling, cell-compatible hydrogels. Its major components Mal-Dextran and the crosslinker CD-Link. CD-Link is composed of polyethylene glycol and a matrix metalloprotease (MMP)-cleavable peptide. The MMP-cleavable peptide is designed for a broad range of MMP cleavage including MMPs MMP1, MMP3, MMP7 and MMP9. It allows cells to spread and migrate within the hydrogel if they express the indicated MMPs.
  • Dextran-PEG Hydrogel SG Kit: a Ready-To-Design kit that provides reagents for setting up slow gelling, cell-compatible hydrogels. Its major components are SG-Dextran and the crosslinker PEG-Link.
  • Dextran-CD Hydrogel SG Kit: a Ready-To-Design kit that provides reagents for setting up slow gelling, cell-compatible hydrogels. Its major components are SG-Dextran and the MMP-cleavable crosslinker CD-Link.
  • Dextran-HA Hydrogel Kit: a Ready-To-Design kit that provides reagents for setting up hydrogels containing hyaluronic acid (hyaluronan). Its major components are SG-Dextran and the thiol-functionalized hyaluronic acid crosslinker HyLink. The hydrogel allows cell spreading and migration if cell adhesion peptides are present in the gel.

Dextranase Enzyme

  • 3-D Life Dextranase: this enzyme allows the degradation of dextran-based 3-D Life Hydrogels. It contains dextranase from Chaetomium sp., which cleaves glycosidic bonds of dextran, thus enabling recovery live or fixed cells without proteolytic degradation.

Cell Recovery Protocol

Ilex Life Sciences LLC is an official distributor of Cellendes GmbH 3-D Life hydrogel products. To learn more about 3-D Life hydrogels, please visit 3-D Life Biomimetic Hydrogels.