VitroGel® HEK293

VitroGel® HEK293 is a xeno-free (animal origin-free) functional hydrogel system developed to support three-dimensional (3D) cultures of human embryonic kidney 293 (HEK293) cells.

• Simple 5-minute protocol

• High-quality 3D spheroids

• Easily scale up with flasks and bioreactors

• Easy cell harvesting protocol (Enzyme-free, No trypsin)

VitroGel HEK293 is a unique system to easily support the rapid 3D growth of HEK293 cells. Cells directly thawed from liquid nitrogen or passaged from 2D culture vessels can be immediately mixed with the hydrogel solution for 3D static suspension cultures. Both our standard 3D cell culture protocol and the 3D static suspension culture protocols can be used for rapid cell expansion. The high-quality 3D spheroids enhance the cell performance for protein expression and downstream applications. HEK293 cells cultured in VitroGel HEK293 can easily be scale-up for large-scale bioreactors or spin flasks. By using the VitroGel Cell Recovery Solution, the cell harvesting after 3D culture is simple and effective.

“Just add cells” 5 min protocol 3D static suspension culture

VitroGel HEK293 is ready-to-use. Just mix with your cells. There is no cross-linking agent or the need to adjust the hydrogel concentration.

Recommended Product

Recover cells from the hydrogel within 15 minutes with high cell viability. Non-enzymatic formulation.

VitroGel® Organoid Recovery Solution

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

VitroGel HEK293 is ready-to-use. Just mix with your cells. There is no cross-linking agent or the need to adjust the hydrogel concentration.

DATA AND REFERENCES

3D Cell Model

Figure 1. 3D static suspension culture of HEK293 cells in VitroGel HEK293.

Cells were mixed with VitroGel HEK293 at (2:1 gel/cells v/v ratio) and then mixed with cell culture medium at 1:3 ratio (cell-gel mixture: culture medium at 1:3 v/v). HEK293 cells form cell spheroids in the hydrogel. The images show the growth and expansion from day 0 to day 7. The enlarged images show the cell spheroids after 7 days of culture. The cell proliferation was tested by Cell Counting Kit-8 (CCK-8) assay. The curve shows the increase of cell number in 14 days culture.

Figure 2. Cell viability of HEK293 cells in 3D suspension culture in VitroGel HEK293.
3D static suspension cultures of HEK293 cells in VitroGel HEK293 were stained with Cyto3DTM Live/Dead Assay Kit (Cat# BM01). The live cells are shown in green color and dead cells are shown in orange color. The images show cells cultured in VitroGel HEK293 hydrogel perform with high cell viability​

Figure 3. 3D static suspension culture of HEK293 cells with high cell density seeding (24 hours).
For protein production purposes, cell suspension can be prepared at high cell density (107 cells/mL) to reach the peak of protein expression within 48 to 72 hours. The images show the fast formation of cell spheroids with over 100 µm size in diameter at 24 hours after cell seeded in VitroGel HEK293 hydrogel for static suspension culture.

Figure 4. Subculture of HEK293 cell spheroids with VitroGel.
The fibroblast-like mouse bone marrow stromal cells (OP9-GFP) were 3D cultured in The cell spheroid culture with VitroGel can be collected by centrifuging (100 x g, 3 min. Please check the cell harvesting protocol of VitroGel Cell Recovery Solution (CAT# MS03-100 for details). The collected cell spheroid can be dissociated into single cells by trypsin. After that, the cells can be resuspended with culture medium and mixed with VitroGel HEK293 for subculture. The images show the formation of cell spheroids from day 0 to day 5 after subculture.

REFERENCES/PUBLICATIONS

• Teryek, M., Jadhav, P., Bento, R., & Biju Parekkadan. (2023). High-Throughput Production of Microcapsules for Human Bone Marrow Derived Mesenchymal Stem Cell Biomanufacturing in a Vertical-Wheel Bioreactor. Biotechnology and Bioprocess Engineering, 28(4), 528–544. https://doi.org/10.1007/s12257-023-0020-9

• Powell K. Adding depth to cell culture. Science, 356(6333), 96–98. https://doi.org/10.1126/science.356.6333.96