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VitroGel 3D is a ready-to-use hydrogel system providing an in-vivo like microenvironment for cell cultures. Cell lines used with the VitroGel 3D system have been used for multiple applications such as drug discovery, toxicity study, cancer biology, stem cell study, and tissue engineering. Scientists are more than satisfied with the user-friendly system and the promising results achieved with the simple operating steps.

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The table below lists examples of cells that have been successfully tested on the VitroGel 3D system. To get the best results from the VitroGel 3D system, the hydrogel conditions can be optimized depending on cell types and components of culture media.  A test of cell growth in a gradient of hydrogel concentrations is recommended for each cell line.

Cell types/applications Product used Culture medium Dilution (v/v) Mixing ratio (v/v)
Hela cells VitroGel 3D-RGD DMEM with 10%FBS 1:3 2:1
MCF-7 breast cancer cells VitroGel 3D-RGD DMEM with 10%FBS 1:3 3:1
HCT-8 Human colon cancer cells VitroGel 3D-RGD RPMI 1640 with 10% FBS 1:3 1:1
Human iPSCs VitroGel 3D-RGD mTeSR1 1:3 3:1
Human embryonic kidney 293 cells VitroGel 3D-RGD DMEM with 10%FBS 1:3 2:1
Beta TC3 cells VitroGel 3D-RGD DMEM 1:3 2:1
Ins-1 cells VitroGel 3D-RGD RPMI 1640 1:3 1:1
BL5 human beta cells VitroGel 3D DMEM with 10%FBS 1:3 2:1
Priess human lymphoblastoid cells VitroGel 3D RPMI 1640 with 10% FBS 1:3 1:3
CD8+ T cells VitroGel 3D RMPI 1640 with 10% FBS 1:3 1:3
Human islets VitroGel 3D-RGD CMRL 1066 medium 1:3 1:3
DLD1 (colon) VitroGel 3D-RGD high glucose DMEM with 10%FBS 1:3 3:1
HepG2 (liver) VitroGel 3D-RGD high glucose DMEM with 10%FBS 1:3 3:1
HuH7 VitroGel 3D-RGD DMEM 1:3 3:1
Prostate cancer line VitroGel 3D-RGD RPMI 1640 1:3 1:1
H460 cells VitroGel 3D-RGD DMEM with 10%FBS 1:3 3:1
A549 cells VitroGel 3D-RGD DMEM with 10%FBS 1:1 4:1
MCF-12A VitroGel 3D-RGD DMEM-F12 1:3 2:1
SKB3 cells VitroGel 3D-RGD DMEM with 10%FBS 1:3 3:1
MDA-MB-231 VitroGel 3D-RGD Mammosphere medium 1:3 1:1
T47D VitroGel 3D-RGD RPMI 1640 with 10% FBS 1:3 1:1
Au565 VitroGel 3D-RGD RPMI 1640 with 10% FBS 1:3 1:1
Human Nthy-ori 3-1 cells VitroGel 3D RPMI 1640 with 10% FBS 1:3 1:1
PDX models of cervix cancer VitroGel 3D Alpha MEM 1:2 4:1
4T1 murine breast cancer cells VitroGel 3D-RGD RPMI 1640 with 10% FBS 1:2 1:1
Melanoma cell lines (M111, M101, and CG-1241) VitroGel 3D-RGD RPMI 1640 1:3 1:1
BT 474 VitroGel 3D-RGD DMEM+10% FBS 1:3 3:1
OVCAR-3 VitroGel 3D-RGD RPMI-1640+10% FBS 1:1 1:1
U87-MG VitroGel 3D-RGD EMEM+10% FBS 1:2 3:1
UD 145 VitroGel 3D-RGD EMEM+10% FBS 1:3 1:1
HCT116 VitroGel 3D-RGD McCoy’s 5A+10% FBS 1:3 1:1
  • The dilution or mixing ratio mentioned above is v/v ratio: for example, pre-gel solution and DI water diluted at 1:3 ratio means 100 μL hydrogel solution mixed with 300 μL DI water.
  • Dilution: The hydrogel solution can be diluted with DI water to adjust the final hydrogel strength. To dilute the VitroGel solution with DI water before mixing with the cell media for gelation, directly mix the solution with DI water or 0.1X PBS at 1:0, 1:1, 1:2, 1:3, 1:4, 1:5 (hydrogel solution : DI water, v/v) ratio at room temperature. (The final gel strength 1:0 > 1:1 > 1:2 > 1:3 > 1:4 > 1:5).
  • Mixing ratio for hydrogel formation: Different mixing ratios of diluted hydrogel solution and cell culture medium would affect the speed of hydrogel formation and the final gel strength. The suggested mixing ratio is from 4:1 to 1:3. (diluted hydrogel solution : cell culture medium, v/v).

Case Studies

VitroGel 3D has been successfully used on different projects with many cell lines. Scientists are impressed by this incredibly easy to use hydrogel system and happy with the promising results. The case studies below show both 3D cell culture and 2D coating of VitroGel 3D system. In each case, all images were taken under the same scale.

Case 1: Beta TC3 cells (used VitroGel 3D-RGD for 3D culture)

Seeding number: 5×105 cells/mL

Day 1

 Day 5

Day 14

Case 2: Ins-1 cells(used VitroGel 3D-RGD for 3D culture)

Seeding number: 5×105 cells/mL

Day 1

Day 5

Day 14

Case 3: Human Foreskin Fibroblast (hFS) derived Induced Pluripotent Stem Cells (iPSCs) (used VitroGel 3D-RGD for both 2D coating and 3D culture)

Seeding number: 2.5×105 cells/mL

hFS iPSCs grown on 2D Matrigel coating (comparison)

Culture on 2D VitroGel 3D-RGD coating (Day 1)

3D culture using VitroGel 3D-RGD (Day 1)

Culture on 2D VitroGel 3D-RGD coating (Day 7)

3D culture using VitroGel 3D-RGD (Day 7)

Summary
  • Both 2D coating and 3D culture are suitable for hFS iPSC culture
  • hFS iPSCs in 2D coating conditions are more defined and rounded in morphology compared to 3D  culture conditions
  • Colonies are rounded as expected within 7 days in hydrogel

Case 4: Beta Lox 5 (BL5) cells (used both VitroGel 3D and VitroGel 3D-RGD for 2D coating and used VitroGel 3D for 3D culture)

Seeding number: 2.5×105 cells/mL

2D cell culture on regular well-plate (control)

Normal human islets grew in suspension (Comparison)

Culture on 2D VitroGel 3D coating for 7 days

3D culture using VitroGel 3D    (Day 1)

Culture on 2D VitroGel 3D-RGD coating for 7 days

3D culture using VitroGel 3D    (Day 7)

Summary
  • BL5 cells grow very well in both 2D coating and 3D culture of VitroGle 3D system
  • Under 2D coating, BL5 cells form colonies and seem to be flat
  • Under 3D culture of VitroGel 3D, BL5 cells form islet-like structures very similar to normal human islets


Case 5: Human Embryonic kidney (HEK293) cells (used VitroGel 3D-RGD for 3D culture)

Seeding number: 2.5×105 cells/mL

2D cell culture on regular well-plate (control)

3D culture using VitroGel 3D-RGD (Day 7)

Summary
  • HEK 293 cells grow in the form of clusters in 3D culture within VitroGel 3D-RGD

Case 6: Human lymphoblastoid Priess cells (used VitroGel 3D for both 2D coating and 3D culture)

Seeding number: 2.5×105 cells/mL

Priess cells grown in suspension (control)

Culture on 2D VitroGel 3D coating for 7 days

3D culture using VitroGel 3D    (Day 7)

Summary
  • Priess cells grown in both 2D coating and 3D culture conditions are similar to cells grown in suspension


Case 7:  CD8+ T cells (use VitroGel 3D for 3D culture)

Seeding number: 2.5×105 cells/mL

CD8+ T cells grew in suspension (control)

3D culture using VitroGel 3D    (Day 7)

Summary
  • CD8+ T cells look vibrant in 3D culture conditions of VitroGel 3D


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Our success stories are growing everyday. Start your cell culture studies with VitroGel 3D today and enjoy your scientific exploration with simplicity and versatility!

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