Overview

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.

Specifications

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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

3D cell culture process in 20 min – “Just add cells”

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.

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Handbooks and Resources

Product Documentation

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• Product Data Sheet v1.0

• Protocol

• Cell Recovery from Gel

• Immunofluoroscence Staining

• Sample Preparation for Histological Analysis

• Cell Fluorescent Staining in Gel for Nuclei and Actin Filaments

• Cell Lysis for Downstream Analysi

Data and References

Xeno-free 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 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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