3D Organoid Culture – Xeno-free Hydrogel

by | Dec 1, 2020 | Organoids, Stem Cell


VitroGel® ORGANOID  are xeno-free (animal origin-free) hydrogels that support the growth of patient-derived organoids or organoids developed from pluripotent stem cells (PSCs).

VitroGel ORGANOID hydrogels are ready-to-use at room temperature and have a neutral pH, transparent, permeable, and compatible with different imaging systems. The solution transforms into a hydrogel matrix by simply mixing with the cell culture medium. VitroGel ORGANOID hydrogels are good for both 3D cell culture and 2D hydrogel coating applications.

The hydrogels can work together with VitroGel STEM (Cat# VHM02), a hydrogel system for 3D static suspension cultures and scale-up of human pluripotent stem cells, by transferring the stem cell spheroids from VitroGel STEM to VitroGel ORGANOID hydrogels for organoid differentiation. The key growth factors and molecules can directly mix with the hydrogel matrix or add on the top of the hydrogel. Organoids cultured in this system can be easily harvested out with our VitroGel Cell Recovery Solution.  VitroGel ORGANOID hydrogels provide a well-defined 3D microenvironment for the future of personalized medicine.

The VitroGel ORGANOID Discovery Kit (Cat# VHM04-K) includes all four types of organoid hydrogels (VitroGel ORGANOID-1, VitroGel ORGANOID-2, VitroGel ORGANOID-3, VitroGel ORGANOID-4) which were formulated with various bio-functional ligands, mechanical strengths, and degradability to fulfill the needs of different organoid culture conditions.

“Just add cells” 5 min protocol. No matrix coating required.

VitroGel ORGANOID is ready-to-use. Just mix with your cells. There is no cross-linking agent or the need to adjust the hydrogel concentration.

VitroGel® ORGANOID (Catalog Number: VHM04-K)

ready-to-use, xeno-free (animal origin-free) hydrogel system for organoid culture


Formulation Xeno-free, functional hydrogel
Use Organoid culture
Operation Ready-to-use at room temperature
Biocompatibility Biocompatible, safe for animal studies
Injection Injectable hydrogel for in vivo studies and lab automation
Cell Harvesting Easy cell recovery using VitroGel Cell Recovery Solution
pH Neutral
Storage Store at 2-8°C. Ships at ambient temperature
Sizes Single Vial: 10 mL
Discovery Kit: (2 mL) each of ORGANOID-1, ORGANOID-2, ORGANOID-3, ORGANOID-4
Usage 60 uses for 2 mL bottle at 50 µL/test
300 uses for 10 mL bottle at 50 µL/test

VitroGel Organoid Methods vs Natural ECM Methods

VitroGel ORGANOID system can culture organoids in variety of methods:

  • 3D cell culture encapsulation
  • 2D hydrogel coating
  • Hydrogel-Cell droplet (A unique method only to VitroGel ORGANOID)

Review all the following VitroGel ORGANOID protocol methods compared to the natural ECM methods and choose the one that best fit s your project

3D Cell Culture Protocol

2D Hydrogel Coating Protocol

Hydrogel-Cell Droplet Protocol

Data and References

Xeno-free 3D Organoid Workflow A: 
Using VitroGel ORGNAOID for organoid culture and maturation

Figure 1. Mouse intestinal organoid culture on VitroGel ORGANOID and Matrigel.
Small organoids recovered from liquid nitrogen were directly seeded with VitroGel and Matrigel, respectively. Images show the growth of mouse intestinal organoid from day 0 to day 14.

Xeno-free 3D Organoid Workflow B:
Start from iPSC spheroids for stem cell differentiation and organoid formation.
(Diagram below shows culturing human intestinal organoid from stem cell spheroids)

Figure 2. Culture human intestinal organoid from stem cell spheroids.
Human iPSCs recovered from liquid nitrogen were seeded with VitroGel STEM for static suspension culture (checking VHM002 for protocols). The high-quality stem cell spheroids formed within 3-5 days with full pluripotent properties (showing the positive markers of SSEA4, OCT4, SOX2, and TRA-1-60). The spheroids were harvested by centrifuging (100g, 3 min) and resuspended with VitroGel STEM in endoderm differentiation medium for 3 days. The endoderm cell spheroids were then harvested by centrifuging (100g, 3 min) and resuspended with VitroGel STEM in mid/hindgut differentiation medium for 3-4 days. The mid/hindguts were collected by centrifuging (100g, 3 min) and characterized with CDX2 and E-Cadherin. Resuspended the mid/hindgut with organoid formation medium and mix with VitroGel ORGANOID (follow the protocol of VitroGel ORGANOID) for organoid formation and long-term maturating culture.

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