VitroGel 3D High Concentration

VitroGel^® 3D High Concentration is a tunable, xeno-free (animal origin-free) hydrogel system that allows the maximum flexibility to manipulate the 3D cell culture environment for different needs. VitroGel 3D High Concentration comes with VitroGel Dilution Solution to adjust the final hydrogel strength from 10 to 4000 Pa. The hydrogel’s tunability gives researchers the ability to create an optimized environment for cell growth. The VitroGel 3D High Concentration hydrogel matrix structure is good for cell spheroid formation, suspension cells, or cells requiring low cell-matrix interactions.

VitroGel High Concentration hydrogels are our xeno-free, tunable hydrogels for researchers wanting full control to manipulate the biophysical and biological properties of the cell culture environment. The tunability of the hydrogel gives the ability to create an optimized environment for cell growth. The hydrogel system has 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. No cross-linking agent is required. Cells cultured in this system can be easily harvested with our VitroGel® Cell Recovery Solution. The hydrogel can also be tuned to be injectable for in vivo studies.

From 3D cell culture, 2D cell coating to animal injection, VitroGel makes it possible to bridge the in vitro and in vivo studies with the same platform system.

Specifications

Contents	VitroGel® 3D High Concentration, 3 mL VitroGel® Dilution Solution, 50 mL
Hydrogel Formulation	Xeno-free tunable hydrogel, pure and unmodified.
Use	Good for cell spheroid formation, suspension cells or cells require low cell-matrix interactions
Mix & Match	Can be blended with other versions of VitroGel concentrated hydrogels to create a custom multi-functional matrix.
Operation	Room temperature
Hydrogel Strength	10 to 4,000 Pa of G’ depending on dilution ratio. Dilute with VitroGel Dilution Solution (TYPE 1 or TYPE 2) for different concentrations.
pH	Neutral
Color	Transparent
Cell Harvesting	VitroGel Organoid Recovery Solution 5-15 min cell recovery
Injectable	Injectable hydrogel
Storage	Store at 2-8°C. Ships at ambient temperature
Number of Uses	Dilution ratio: 1:2 = 225 uses at 50 µL per well 1:3 = 300 uses at 50 µL per well 1:5 = 450 uses at 50 µL per well

3D Cell Culture Process in 20 Minutes

VitroGel High Concentration hydrogels are easy-to-use. There is no cross-linking agent required. Work confidently at room temperature.

Tunable Hydrogel Strength

Simply diluting the hydrogel controls the gel strength

Handbooks and Resources

Protocol/Handbooks

 VitroGel 3D Protocol
  Cell Recovery/Harvesting Protocol
 Live-Dead Cell Viability Assay Protocol
 Injection Protocol
  Five Cell Culture Methods for VitroGel

  Cell Fluorescent Staining in Gel for Nuclei
and Actin Filament
RNA/DNA Extraction, Cell Lysis Protocol for Downstream Analysis
  Cell Proliferation Assay
  Immunofluorescence Staining
  Sample Preparation for Histological Analysis

Product Documentations

Frequently Asked Questions
 Sales Sheet
 Product Data Sheet
 Material Safety Data Sheet (MSDS)

Video Protocols & Demonstrations

VIDEO PROTOCOL TIP

3D Cell Culture
VitroGel High Concentration Hydrogels

VIDEO PROTOCOL TIP

2D Cell Coating
Method 1
VitroGel High Concentration Hydrogels

VIDEO PROTOCOL TIP

2D Cell Coating
Method 2
VitroGel High Concentration Hydrogels

VIDEO PROTOCOL TIP

Cell Recovery
Method 1
from 3D & 2D cultures

VIDEO PROTOCOL TIP

Cell Recovery
Method 2
from 3D & 2D  cultures

VIDEO PROTOCOL TIP

Injectable Hydrogel
VitroGel System

Application Notes

APPLICATION NOTE

Long-Term Neuron Culture Maturation in 3D Hydrogel Constructs

APPLICATION NOTE

3D Cell Culture of Human Colon Cancer Cells (HCT116) on VitroGel® System

APPLICATION NOTE

3D Cell Culture of Human Pancreatic Cancer Cells (PANC-1) on VitroGel® System

APPLICATION NOTE

3D Invasion of Glioblastoma Cells in VitroGel Hydrogel System

Research Highlights

RESEARCH HIGHLIGHT

Long-Lived Lab Grown Cells for Diabetes Treatment

RESEARCH HIGHLIGHT

Finding Living Needles in a 3D Haystack

RESEARCH HIGHLIGHT

Cutting Colon Cancer Out Properly

RESEARCH HIGHLIGHT

Go With the PVP Flow for Invitro Model of Hydrocephalus

RESEARCH HIGHLIGHT

No Relaxing in the Fight Against Liver Cancer

Data and References

Cell Type Behavior Reference Table – VitroGel 3D

Studies 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 culture on the surface of regular tissue culture treated well plate (control); B. Normal human islets grew 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 the 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.

References/Publications

• References to all VitroGel hydrogels >
• Castañeyra-Ruiz, L., Lee, S., Chan, A. Y., Shah, V., Romero, B., Ledbetter, J., & Muhonen, M. (2022). Polyvinylpyrrolidone-Coated Catheters Decrease Astrocyte Adhesion and Improve Flow/Pressure Performance in an Invitro Model of Hydrocephalus. Children, 10(1), 18. https://doi.org/10.3390/children10010018
  
• Wei, J., Yao, J., Yang, C., Mao, Y., Zhu, D., Xie, Y., Liu, P., Yan, M., Ren, L., Lin, Y., Zheng, Q., & Li, X. (2022). Heterogeneous matrix stiffness regulates the cancer stem-like cell phenotype in hepatocellular carcinoma. Journal of Translational Medicine, 20(1). https://doi.org/10.1186/s12967-022-03778-w
  
• Yu, Y., Wu, X., Wang, M., Liu, W., Zhang, L., Zhang, Y., Hu, Z., Zhou, X., Jiang, W., Zou, Q., Cai, F., & Ye, H. (2022). Optogenetic-controlled immunotherapeutic designer cells for post-surgical cancer immunotherapy. Nature Communications, 13(1), 6357. https://doi.org/10.1038/s41467-022-33891-9
  
• Manferdini, C., et al. (2022). RGD-Functionalized Hydrogel Supports the Chondrogenic Commitment of Adipose Mesenchymal Stromal Cells Gels. https://www.mdpi.com/2310-2861/8/6/382
• Ouyang,L., et al.(2022) Overexpressing HPGDS in adipose-derived mesenchymal stem cells reduces inflammatory state and improves wound healing in type 2 diabetic mice. Stem Cell Research & Therapy, 2022,13:395. https://stemcellres.biomedcentral.com/articles/10.1186/s13287-022-03082-w
• Worden, Austin N.(2022) A novel model to study adipose-derived stem cell differentiation. University of South Carolina ProQuest Dissertations Publishing, 2022, 28967872. https://www.proquest.com/openview/726a089f8f0894146e3f9bf083913e3a/1?pq-origsite=gscholar&cbl=18750&diss=y
• Fen, et al.(2022) Optimization of Three-Dimensional Culture Conditions of HepG2 Cells with Response Surface Methodology Based on the VitroGel System. Biomedical and Environmental Sciences, 2022,(35,8), 688-698. https://www.frontiersin.org/articles/10.3389/fimmu.2022.914381/full
• Yamazaki et al.(2022) Assessment of hypoxia-targeting therapy for intestinal T-cell lymphoma in dogs: preclinical test using murine models. Available at SSRN: https://ssrn.com/abstract=4090297 or http://dx.doi.org/10.2139/ssrn.4090297
• Cui, J., et al. (2022). ATR inhibition sensitizes liposarcoma to doxorubicin by increasing DNA damage. American Journal of Cancer Research. https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9077062/
• Fengyuan, M. Y., Shen, L., Fan, D. D., Bai, Y., Li, B., & Lee, J. (2022). YAP9/A20 complex suppresses proinflammatory responses and provides novel anti-inflammatory therapeutic potentials. Frontiers in Immunology. https://www.frontiersin.org/articles/10.3389/fimmu.2022.914381/full
  
• Sinjushin, A., et al. (2022). Variations in Structure among Androecia and Floral Nectaries in the Inverted Repeat-Lacking Clade (Leguminosae: Papilionoideae) Plants, Special Issue: Floral Secretory Tissue: Nectaries and Osmophores. https://www.mdpi.com/2223-7747/11/5/649
• Worden, A., et al. (2022). Self-Assembling Toroidal Cell Constructs for Tissue Engineering Applications Microscopy and Microanalysis. https://doi.org/10.1017/S1431927622000253
• Chen, Y., et al. (2021). Ultra-sensitive responsive near-infrared fluorescent nitroreductase probe with strong specificity for imaging tumor and detecting the invasiveness of tumor cells Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy. https://doi.org/10.1016/j.saa.2021.120634
• Powell K.(2017) Adding depth to cell culture. Science, 356(6333), 96–98. https://doi.org/10.1126/science.356.6333.96