Protocols and Resources

VitroGel® 3D Protocol

3D/2D Culture Protocol Quick Guide

Xenograft/Injection Protocol

Five Cell Culture Methods for VitroGel® Protocol

Cell Recovery/Harvesting Protocol

Live-Dead Cell Viability Assay Protocol

Cell Fluorescent Staining in Gel for Nuclei and Actin Filament Protocol

RNA/DNA Extraction, Cell Lysis for Downstream Analysis Protocol

Cell Proliferation Assay Protocol

Immunofluorescence Staining Protocol

Sample Preparation for Histological Analysis Protocol

Product Documentation

Sales Sheet – VitroGel® High Concentration

 Sale sheet – Injectable synthetic hydrogel for excellent delivery

Product Data Sheet

Material Safety Data Sheet (MSDS)

Frequently Asked Questions 

Data and References

Cell Type Behavior Reference Table – VitroGel® 3D

Studies were performed using VitroGel® 3D in different tissue and cell types.

Data

Figure 1. Beta Lox 5 (BL5) cells 3D culture in VitroGel® 3D system.
A. BL5 cells cultured on the surface of a regular tissue culture-treated well plate (control); B. Normal human islets grown in suspension culture (comparison); C. 3D culture of BL5 cells in VitroGel 3D at Day 1; D. 3D culture of BL5 cells in VitroGel 3D at Day 7. Under 3D culture of VitroGel 3D, BL5 cells form islet-like structures very similar to normal human islets. The hydrogel is prepared at 1:3 dilution. The images were taken at 10X magnification.

Figure 2. CD8+ T cells 3D culture in VitroGel® 3D system.
CD8+ T cells culture grew in suspension culture (control); B. 3D culture of CD8+ T cells in VitroGel 3D at Day 7.  CD8+ T cells are vibrant in 3D culture conditions of VitroGel 3D. The cells can easily move within the unmodified hydrogel matrix. The hydrogel is prepared at 1:3 dilution. The images were taken at 10X magnification.

2D Coating Applications

Figure 3. Human colon cancer cells (HCT 116) cells cultured on top of VitroGel® 3D hydrogel.
A thick hydrogel coating plate has been prepared by mixing VitroGel® 3D with PBS at 1:1 ratio. A 300 µL mixture has been added to a well of a 24-well plate and stabilization at room temperature for 20 minutes before adding cells on top of the hydrogel. Cell spheroids form on top of the hydrogel. Cells seeded at 2.5-10×10⁵ cells/mL.

Figure 4. Comparison of long-term neuronal culture seeded onto thick hydrogel mats.
Cells are stained with Beta-III-Tubulin (green) cytoskeleton marker and their nuclei are counter-stained with DAPI (blue). Cells spread out and form neural-like networks as early as day 3 post-differentiation, with comparable efficacy between VitroGel 3D and Matrigel, based on cell survival, culture spreading, and morphological analysis, reached between days 7 and 9. On Matrigel mats, cell culture health and viability drop off sharply once day 9 has passed, with most cells detaching and neurites retracting by day 14, and the vast majority of cells gone by day 21. If grown onto VitroGel® 3D mats, differentiated B35 neurons have a tendency to self-organize into 3D clusters very early on (Day 7), assuming a mixed 2D/3D cell culture for the first two weeks of the time course. By Day 21, these cells have migrated into self-assembled 3D clusters, embedded into the thick hydrogel matrix, with very few cells between the clusters, but without any significant cell death.

Figure 5. Human Lymphoblastoid Priess cells cultured on top of VitroGel® 3D hydrogel.
A. Priess cells grown in suspension (control); B. Priess cells grown on top of VitroGel® 3D at day 7. A hydrogel substance can be prepared with different stiffness by adjusting the dilution of VitroGel 3D from 1:1 to 1:3 ratio. Cells seeded on the top of the hydrogel form cell spheroids form on the top of the hydrogel. The hydrogel provides a soft substance for cells to attach and grow.

Video Protocols & Demonstrations

Application Notes

Research Highlights

References/Publications

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