VitroGel® 3D High Concentration

tunable, xeno-free hydrogel, high concentration

  • 3 mL Kit
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SKU: TWG001 Category:

Overview

VitroGel® 3D High Concentration is a tunable, xeno-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 tunability of the hydrogel gives the ability for researchers 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

ContentsVitroGel® 3D High Concentration, 3 mL
VitroGel® Dilution Solution, 50 mL
Hydrogel FormulationXeno-free tunable hydrogel, pure and unmodified.
HIGH CONCENTRATION
UseGood for cell spheroid formation, suspension cells or cells require low cell-matrix interactions
OperationRoom temperature
Hydrogel Strength10 to 4,000 Pa of G’ depending on dilution ratio.
Dilute with VitroGel Dilution Solution (TYPE 1 or TYPE 2) for different concentrations.
pHNeutral
ColorTransparent
Cell Harvesting20 min cell recovery using VitroGel Cell Recovery Solution
InjectableInjectable hydrogel
StorageStore at 2-8°C. Ships at ambient temperature

3D cell culture process in 20 min

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

Product Documentation

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Technical Data Sheet

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Frequently Asked Questions

View FAQ (Expand)

To see a full list of FAQ, click here.  FAQ LIST

  • How to prepare the cell suspension to mix the hydrogel? Shall I add serum?
    If cells cultured in a complete cell culture medium, which is supplement with 10% FBS or other critical supplements, please prepare the cell suspension using the following methods before mixing it with hydrogel solution.
  1. Prepare the cell suspension with 2X concentration (e.g. 100K), and mix with 100% FBS at 1:1 (v/v) ratio to get 1X cell suspension (50K) with 50% FBS.
  2. Mix the diluted hydrogel solution with the cell suspension from above at 4:1 (v/v) ratio to get the final cells in the hydrogel at 10K with 10% FBS supplement.

If serum plan is an important role in your traditional cell culture, it is also important for 2D coating and 3D culture. Adding serum supplements in the hydrogel and adjusting the final serum concentration to the target level would support cell growth in the hydrogel system.

  • How do I adjust the hydrogel formation time?

    –  If VitroGel needs to be diluted more than 1:3 ratio, a longer waiting time (20-30 min) may be needed for soft gel formation. Using a higher volume of cell culture medium for mixing would help to accelerate the process of hydrogel formation.-  If the hydrogel solidifies too fast after mixing with culture medium (showing as small solid gel chunk), adjust the mixing ratio by using less cell culture medium. For example, if mixing 4 mL diluted hydrogel solution with 1 mL cell culture medium lead to the solid gel chuck (particles), then mixing 4 mL diluted hydrogel solution with 0.5-0.8 mL cell culture medium would help to solve the issue.-  On the other hand, if the hydrogel formation is too slow, which may happen when using low hydrogel concentration at 1:3 or 1:4 dilution or using a cell culture medium with very low ionic concentration, adjust the mixing ratio by using more cell culture medium. For example, if mixing 4 mL diluted hydrogel solution with 1 mL cell culture medium lead to a slow hydrogel formation, then mixing 4 mL diluted hydrogel solution with 1.5-4 mL cell culture medium would help to solve the issue.
  •  How do I adjust the stiffness of the final hydrogel?
    The stiffness of the final hydrogel can be adjusted by diluting the hydrogel solution before mixing with cell culture media. Our VitroGel Dilution Solution can help to adjust the hydrogel concentration. Please read the “First-time User Note” to learn how to prepare different VitroGel dilutions. If you need a higher hydrogel stiffness than the original product, please contact us at support@thewellbio.com.

  • Can I harvest cells from the hydrogel after 3D culture?
    Yes, the cells can be harvested after 2D coating or 3D culture by using the VitroGel Cell Recovery Solution. VitroGel™ Cell Recovery Solution is a ready-to-use, enzyme-free solution to harvest 2D or 3D cultured cells from hydrogel fast and safely. The solution is compatible with the VitroGel hydrogel system and can recover cells from VitroGel in 15 minutes. VitroGel Cell Recovery Solution is room temperature stable, has a neutral pH and works at 37 °C operating temperature. The solution can maintain high cell viability during the recovery process. Cells can be sub-culture in both 2D and 3D culture after recovery.

 

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Resources

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Application Notes

APPLICATION NOTE

Data and References

Cell Type Behavior Reference Table for VitroGel 3D

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

Cancer/Tumor
Cell TypeBehavior
Brainstem glioma DIPGCell proliferation and survival
Breast CTCCell proliferation
Breast T47DSpheroid formation and proliferation
Chordoma CellsCell proliferation
Hela CellsCell proliferation
Human osteosarcoma KHOSCell proliferation and spheroid formation
Human osteosarcoma U2OSCell proliferation and spheroid formation
Epithelial Cells
Cell TypeBehavior
Human NTHY-ORI 3-1 CellsEnhance spheroids and cluster formation and promote cell viability
Immune Cells
Cell TypeBehavior
BL5 human beta cellsEnhance spheroids and cluster formation and promote cell viability
CD8 + T cellsEnhance spheroids and cluster formation and promote cell viability
Priess human lymphoblastoid cellsEnhance spheroids and cluster formation and promote cell viability
Red Blood Cells
Cell TypeBehavior
Red Blood CellsEnhance spheroids and cluster formation and promote cell viability
Stem Cells
Cell TypeBehavior
Human stem cells from apical papilla SCAPEnhance cell viability
All Cell Type Behaviors for All VitroGel Products
Tissue/Organ typeCell TypeRelate productBehavior
Beta cellBL5 human beta cellsVitroGel Hydrogel Matrix, VitroGel 3DEnhance spheroids and cluster formation and promote cell viability.
Beta TC3 cellsVitroGel Hydrogel Matrix, VitroGel RGDCell proliferation and cellular interations
BoneBone marrow stromal cells (rat)VitroGel Hydrogel Matrix, VitroGel RGDOsteogenesic differentiation
VitroGel Hydrogel Matrix, VitroGel RGDCell proliferation, cell viability, and cellular networking
VitroGel Hydrogel Matrix, VitroGel COLCell attachment and osteoblast differentiation
Bone marrow mesenchymal stem cells (human)VitroGel Hydrogel Matrix, VitroGel COLChondrogenic/osteogenic differentiation
VitroGel Hydrogel Matrix, VitroGel IKVAVAngiogenesis
VitroGel Hydrogel Matrix, VitroGel COLCell spreading, proliferation, and collagen II production
Bone marrow mesenchymal stem cells (goat)VitroGel Hydrogel Matrix, VitroGel RGDOsteogenesic differentiation
Osteoblasts (rat)VitroGel Hydrogel Matrix, VitroGel RGDCell attachment and spreading
Bone marrow stromal cells (bovine)VitroGel Hydrogel Matrix, VitroGel COLCell spreading and osteocalcin expression
BreastMammary gland MCF10AVitroGel Hydrogel Matrix, VitroGel MMPMMP activity in response to TGF-ß1
Mammary epithelium (mouse)VitroGel Hydrogel Matrix, VitroGel COLCell invasion and dissemination
Cancer/tumorHuman colorectal carcinoma HCT 116VitroGel Hydrogel Matrix, VitroGel RGDcell proliferation, cell survival, and intercelluar networking
Huaman colon carcinoma HCT-8VitroGel Hydrogel Matrix, VitroGel RGDCell proliferation and cell matirx interaction
Glioma U87-MGVitroGel Hydrogel Matrix, VitroGel RGDCell spreading and actin stress fiber assembly
VitroGel Hydrogel Matrix, VitroGel RGDCell proliferation and cell matirx interaction
VitroGel Hydrogel Matrix, VitroGel COLCell migration dependent on mechanical force
VitroGel Hydrogel Matrix, VitroGel MMPcell proliferation, spreading, and migration
Primary glioblastom U87VitroGel Hydrogel Matrix, VitroGel RGDcell proliferation and cellular interations
Glioblastoma SF 268VitroGel Hydrogel Matrix, VitroGel RGDCell proliferation and cell matirx interaction
Glioblastoma SF 295VitroGel Hydrogel Matrix, VitroGel RGDCell proliferation and cell matirx interaction
Glioblastoma SNB75VitroGel Hydrogel Matrix, VitroGel RGDCell proliferation and cell matirx interaction
Glioblastoma U-251 MGVitroGel Hydrogel Matrix, VitroGel RGDCell proliferation and cell matirx interaction
Prostate PC3VitroGel Hydrogel Matrix, VitroGel COLCell proliferation and reduced MMP release
VitroGel Hydrogel Matrix, VitroGel IKVAVcell proliferation and invasion
VitroGel Hydrogel Matrix, VitroGel RGDCell proliferation and invasion
VitroGel Hydrogel Matrix, VitroGel COLCell invasion, migration, and spheroid metabolic activity
Prostate LNCaPVitroGel Hydrogel Matrix, VitroGel RGDCell attachment
VitroGel Hydrogel Matrix, VitroGel COLCell proliferation and prostate specific antigen release
Prostate CRPCVitroGel Hydrogel Matrix, VitroGel RGDCell proliferatin and invasion
Prostate DU145VitroGel Hydrogel Matrix, VitroGel RGDCell proliferation and invasion
Melanoma B16F10VitroGel Hydrogel Matrix, VitroGel COLCell migration, invasion, and MMP release
VitroGel Hydrogel Matrix, VitroGel YIGSRCell attachment and spreading
Breast MDA-MB-231VitroGel Hydrogel Matrix, VitroGel MMPCell invasion
VitroGel Hydrogel MatrixCell spreading
VitroGel Hydrogel Matrix, VitroGel RGDCell proliferation, division, migration, and invasion
VitroGel Hydrogel Matrix, VitroGel COLCell spreading and cluster growth
Fibrosarcoma HT1080VitroGel Hydrogel Matrix, VitroGel COLCell infiltration
VitroGel Hydrogel Matrix, VitroGel COLCell attachment
Breast T47D VitroGel Hydrogel Matrix, VitroGel COLForce dependent tubule formation
VitroGel Hydrogel MatrixCell cluster growth
VitroGel Hydrogel Matrix, VitroGel 3DSpheroid formation and proliferation
VitroGel Hydrogel Matrix, VitroGel COLCell cluster growth
Breast 4T1VitroGel Hydrogel Matrix, VitroGel RGDCell proliferation
Breast CTCVitroGel Hydrogel Matrix, VitroGel 3DCell proliferation
Breast E0771VitroGel Hydrogel Matrix, VitroGel RGDCell proliferation, spheroid formation
Breast AU-565VitroGel Hydrogel Matrix, VitroGel RGDCell proliferation, cell matrix interations
Epithelial ovarian OV-MZ-6VitroGel Hydrogel Matrix, VitroGel RGDSpheroid formation and proliferation
Epithelial ovarian SKOV-3VitroGel Hydrogel Matrix, VitroGel RGDSpheroid formation and proliferation
Glioma U373-MGVitroGel Hydrogel Matrix, VitroGel RGDCell adhesion and migration
Rhabdomyosarcoma (human)VitroGel Hydrogel Matrix, VitroGel YIGSRCell attachment and spreading
Melanoma SK-MEL-28VitroGel Hydrogel Matrix, VitroGel IKVAVCell adhesion and proliferation
Melanoma K-1735VitroGel Hydrogel Matrix, VitroGel IKVAVCell invasion
Melanoma A2058VitroGel Hydrogel Matrix, VitroGel IKVAVCollagenolytic activity
Brainstem glioma DIPGVitroGel Hydrogel Matrix, VitroGel 3DCell proliferation and survival
Hela CellsVitroGel Hydrogel Matrix, VitroGel 3DCell proliferation
Colorectal adenocarcinoma DLD-1 cellsVitroGel Hydrogel Matrix, VitroGel RGDCell proliferation and cell matirx interaction
Glioma LRM55VitroGel Hydrogel Matrix, VitroGel IKVAVCell attachment
Melanoma WM239AVitroGel Hydrogel Matrix, VitroGel MMPCell invasion
Melanoma CellsVitroGel Hydrogel Matrix, VitroGel RGDCell proliferation and cell matirx interaction
Insulinoma ins-1 (Rat)VitroGel Hydrogel Matrix, VitroGel RGDCell proliferation and cell matirx interaction
HEK 293VitroGel Hydrogel Matrix, VitroGel RGDCell proliferation and cell matirx interaction
Biphasic synovial sarcoma SYO-1VitroGel Hydrogel Matrix, VitroGel RGDCell proliferation, cell matirx interaction, and cell survival
Fuji CellsVitroGel Hydrogel Matrix, VitroGel RGDCell proliferation and cell matirx interaction
Chordoma CellsVitroGel Hydrogel Matrix, VitroGel 3DCell proliferation
Bone OSA 1777VitroGel Hydrogel Matrix, VitroGel RGDspheroid and cluster formation
Glioma RuGliVitroGel Hydrogel Matrix, VitroGel COLIntegrin dependent cell adhesion
Breast Cancer MCF-7VitroGel Hydrogel Matrix, VitroGel RGDCell proliferation, intercellular connections
VitroGel Hydrogel Matrix, VitroGel COLCell proliferation, morphological changes, MMP expression, and angiogenesis
Liver carcinoma HepG2VitroGel Hydrogel Matrix, VitroGel COLCell viability, growth, and drug resistance
VitroGel Hydrogel Matrix, VitroGel RGDCell proliferation and cell matirx interaction
Human pancreatic cancer PANC-1VitroGel Hydrogel Matrix, VitroGel RGDcell proliferation and cellular interations
Primary breast (human)VitroGel Hydrogel Matrix, VitroGel COLCell invasion, migration, and dissemination
Ovarian carcinoma OVCAR-3VitroGel Hydrogel Matrix, VitroGel RGDCell proliferation, cell matrix interations
Ovarian OVCA429VitroGel Hydrogel Matrix, VitroGel COLMMP dependent cell invasion
Human osteosarcoma KHOSVitroGel Hydrogel Matrix, VitroGel 3Dcell proliferation and spheroids formation
Human osteosarcoma U2OSVitroGel Hydrogel Matrix, VitroGel 3Dcell proliferation and spheroids formation
Priess human lymphoblastoid cellsVitroGel Hydrogel Matrix, VitroGel 3DEnhance spheroids and cluster formation and promote cell viability.
CartilageChondrocytes (bovine)VitroGel Hydrogel Matrix, VitroGel RGDCell viability and proliferation
Chondrocytes (human)VitroGel Hydrogel Matrix, VitroGel RGDCell viability and proliferation
Connective tissueDermal fibroblasts (human)VitroGel Hydrogel Matrix, VitroGel RGDCell viability and spreading
VitroGel Hydrogel Matrix, VitroGel COLCell viability
Fibroblasts NIH3T3VitroGel Hydrogel Matrix, VitroGel RGDDirectional cell migration toward gradient
VitroGel Hydrogel Matrix, VitroGel COLCell spreading dependent on substrata rigidity
Foreskin fibroblasts (human)VitroGel Hydrogel Matrix, VitroGel RGDCell spreading
VitroGel Hydrogel Matrix, VitroGel YIGSRCell spreading
VitroGel Hydrogel Matrix, VitroGel MMPSubstrata degradation and cell invasion
Skin fibroblasts (skin)VitroGel Hydrogel Matrix, VitroGel IKVAVCell adhesion
Epidermal keratinocytesVitroGel Hydrogel Matrix, VitroGel COLCell viability
Epithelial CellsMouse ovarian follicle cellsVitroGel Hydrogel Matrix, VitroGel RGD3D cell culture using ES-hydrogel can enhance vitro follicle culture by considering the permeability and stiffness of the gel.
Human Nthy-ori 3-1 cellsVitroGel Hydrogel Matrix, VitroGel 3DEnhance spheroids and cluster formation and promote cell viability.
A549 cellsVitroGel Hydrogel Matrix, VitroGel RGDEnhance cell proliferation and cell matrix interactions.
MCF-12AVitroGel Hydrogel Matrix, VitroGel RGDEnhance cell proliferation and cell matrix interactions.
Immortalized bronchial epithelial cells HBEC-KRASVitroGel Hydrogel Matrix, VitroGel 3DCell proliferation
EyeCorneal endothelial B4G12VitroGel Hydrogel Matrix, VitroGel COLCell attachment and spreading
Retinal ganglion cells (xenopus)VitroGel Hydrogel Matrix, VitroGel COLNeurite outgrowth
Immune CellsCD8 + T cellsVitroGel Hydrogel Matrix, VitroGel 3DEnhance spheroids and cluster formation and promote cell viability.
KidneyHuman embryonic kidney HEK293VitroGel Hydrogel Matrix, VitroGel RGD3D spheroids formation
VitroGel Hydrogel Matrix, VitroGel COLCell proliferation and cluster growth
Madin-Darby Canine KidneyVitroGel Hydrogel Matrix, VitroGel RGDEpithelial cysts formation
podocytes (human)VitroGel Hydrogel Matrix, VitroGel COLGlomerular capillary formation
glomerular endothelial cells (human)VitroGel Hydrogel Matrix, VitroGel COLGlomerular capillary formation
LiverHepatocytes (human)VitroGel Hydrogel Matrix, VitroGel RGDFilopodia formation and synthesis of albumin
VitroGel Hydrogel Matrix, VitroGel COLCell attachment
Hepatocytes (mouse)VitroGel Hydrogel Matrix, VitroGel RGDCell viability
Hepatocytes (rat)VitroGel Hydrogel Matrix, VitroGel COLAlbumin secretion
Hepatocytes (swine)VitroGel Hydrogel Matrix, VitroGel COLCell spreading and albumin section
LungAlveolar basal epithelial A549VitroGel Hydrogel Matrix, VitroGel RGDCell attachment
Alveolar epithelial RLE-6TNVitroGel Hydrogel Matrix, VitroGel RGDCell attachment and mesenchymal differentiation
Pulmonary fibroblasts LL2VitroGel Hydrogel Matrix, VitroGel IKVAVCell adhesion
HFL1 lung fibroblasts CCL153VitroGel Hydrogel Matrix, VitroGel COLCell proliferation and spindle morphology
Lung cancer associated fibroblasts (human)VitroGel Hydrogel Matrix, VitroGel COLSubstrata contractility
Lung fibroblasts MCR-5VitroGel Hydrogel Matrix, VitroGel COLNGF-mediated substrata contraction
MuscleMyoblasts C2C12VitroGel Hydrogel Matrix, VitroGel RGDCell Proliferation and differentiation
VitroGel Hydrogel Matrix, VitroGel COLCell attachment, proliferation, and myofibril formation
VitroGel Hydrogel MatrixMyotube formation
VitroGel Hydrogel Matrix, VitroGel COLIntegrin dependent cell adhesion
Skeletal myoblasts (mouse)VitroGel Hydrogel Matrix, VitroGel RGDCell attachment, proliferation, and myofibril formation
Myoblasts (human)VitroGel Hydrogel Matrix, VitroGel COLCell adhesion, alignment along fiber, and myotube formation
Myoblasts C25Cl48VitroGel Hydrogel Matrix, VitroGel COLCell proliferation, differentiation and myotube formation
NeuralDorsal root ganglion (chick)VitroGel RGDNeurite formation and outgrowth
VitroGel COLForce dependent neurite outgrowth
Neural PC12VitroGel COLNeurite outgrowth
VitroGel IKVAVNeurite outgrowth
Neural stem cell/progenitor cell (rat)VitroGel YIGSRCell viability
VitroGel IKVAVCell attachment and differentiation
Neural stem cell/progenitor cell (human)VitroGel IKVAVCell viability and differentiation
VitroGel LDP1Cell viability and differentiation
VitroGel LDP1Cell viability
VitroGel COLCell attachment
Schwann cells (rat)VitroGel YIGSRCell attachment and migration
Neural stem cell/progenitor cell (mouse)VitroGel IKVAVCell adhesion and differentiation
Cortical astrocytes (rat)VitroGel IKVAVCell adhesion
Spiral ganglion neurons (mouse)VitroGel IKVAVNeurite outgrowth
Motor neurons (human)VitroGel COLForce dependent neurite outgrowth
Forebrain neurons (human)VitroGel COLForce dependent neurite outgrowth
Cortical neurons (rat)VitroGel COLNeuronal viability and neurite outgrowth
Dorsal root ganglion (rat)VitroGel COLNeurite outgrowth
Red Blood CellsRed Blood cellsVitroGel Hydrogel Matrix, VitroGel 3DEnhance spheroids and cluster formation and promote cell viability.
PancreasB-cells MIN6VitroGel Hydrogel Matrix, VitroGel IKVAVReduced apoptosis and increased insulin release
Stem cellsMesenchymal stem cells (human)VitroGel RGDCell viability
VitroGel RGDCell Proliferation and differentiation
VitroGel COLCell proliferation
VitroGel IKVAVNeuronal differentiation
VitroGel MMPNeuronal differentiation and neurite outgrowth
VitroGel COLCell attachment, spreading, viability, and osteoblast differentiation
Mesenchymal stem cells (mouse)VitroGel RGDCell spreading and migration
VitroGel MMPCell spreading and migration
Mesenchymal stem cells (rat)VitroGel RGDCell adhesion and spreading
Embryonic stem cells (mouse)VitroGel RGDEndothelial cell differentiation
VitroGel COLNeuronal differentiation and neurite outgrowth
VitroGel YIGSRNeuronal differentiation
Induced pluripotent stem cells (human)VitroGel YIGSRCell viability
VitroGel IKVAVCell viability
VitroGel LDP1Cell viability
Human IpscVitroGel RGDCell proliferation, and cell matrix interactions
Human stem cells from apical papilla SCAPVitroGel 3DCell viability
Adipose derived stem cells (human)VitroGel IKVAVCell attachment
Vascular/cardiacUmbilical vein endothelial cells (human)VitroGel RGDCell attachment, proliferation, migration, and angiogenesis
VitroGel YIGSRUpregulation in gene expression
VitroGel IKVAVMigratory cell infiltration
VitroGel MMPCell attachment, migration, and survival
VitroGel COLCell attachment, spreading, and VEGF dependent migration
Neonatal cardiac (rat)VitroGel RGDCell attachment and tissue regeneration
VitroGel YIGSRCell attachment similar to laminin
Aortic smooth muscle cells (human)VitroGel RGDCell attachment
Endothelial (human)VitroGel YIGSRCell differentiation
EndotheliocytesVitroGel YIGSRCell migration
Microvascular endothelial cells (human)VitroGel YIGSRCell mobility
Aortic endothelial cells (bovine)VitroGel COLForce dependent cell spreading
Capillary endothelial cells (bovine)VitroGel COLCapillary like network formation

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 prepare 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 drops 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 hydrogel. The hydrogel provides a soft substance for cell to attach and grow.

References/Publications

Size

3 mL Kit