VitroGel® STEM

• Establish stem cell spheroids directly from liquid nitrogen (5 min protocol)

• High-quality 3D stem cells for differentiation and organoid formation

• Easy scale-up (from laboratory scale to industrial scale)

• Efficient cell harvesting or subculturing

VitroGel® STEM is a xeno-free (animal origin-free) hydrogel system developed to improve the performance of three-dimensional (3D) static suspension cultures and scale-up of human pluripotent stem cells (hPSCs) to create a high-throughput system to model various tissue and disease states.

This hydrogel system is ready to use with an optimized formulation that fully supports the rapid expansion of high-quality 3D stem cell spheroids with pluripotent properties. hPSCs directly thawed from liquid nitrogen or passaged from 2D matrix coated culture vessels can be immediately mixed with the hydrogel solution for static suspension cultures. Moreover, the optimization protocol is ideal for time-sensitive experiments, as it does not require excessive medium exchanges, which can ultimately save on time and materials. This hydrogel system is compatible with most hPSC culture media and tissue culture vessels. Due to the unique static suspension culture procedure, the requirement for microcarriers for large-scale bioreactors is eliminated, making cell harvesting simple and effective. The 3D stem cell spheroids that are developed using this system can be used for further sub-cultures, patterned differentiations, organoid development, or re-establishing 2D culture morphologies.

“Just add cells” 5 min protocol. No matrix coating required.

VitroGel STEM is ready-to-use. Just mix with your hPSCs. There is no laborious matrix coating required to maintain and expand your stem cells.

Recommended Product

Harvest organoids from the hydrogel in 5-15 minutes with high cell viability. Non-enzymatic formulation.

VitroGel® Organoid Recovery Solution

Advantages of the VitroGel STEM

Easy-to-use

• Easily seed stem cells directly from liquid N2 or 2D culture systems

• Efficient cell harvesting or sub culturing

High-performance

• Can be used for stem cell scale-up with bioreactors at ultra-low agitating speed to maintain high cell viability and excellent cell growth rates

• Yields high-quality 3D stem cells with full pluripotent properties

Flexible

• Undifferentiated stem cells can be easily transferred to 96 well plates, T-flasks, shaking flasks, and bioreactors, making them compatible with most culture vessels

• Compatible with the majority of stem cell culture media

• Easily switched back to 2D culture systems if needed

Cost-effective

• Eliminates the need for matrix coating or microcarriers

• Eliminates the need for shakers or stirrers for laboratory scale stem cell suspension cultures

DATA AND REFERENCES

Figure 1. 3D static suspension culture of hPSC from 2D matrix culture

After 24 hours, small hPSC spheroids start to form. From day 1 to 6, cells in the suspension cultures quickly grow, leading to the generation of healthy and high-quality stem cell spheroids. After day 3, cell numbers grew exponentially (Figure 1B), and spheroid size steadily increases (Figure 1C). The hPSC spheroids display characteristics of shallow craters or pockmarks, indicating expression of hPSC markers and successful expansion of healthy and high-quality stem cell spheroids. The resulting spheroids provide researchers with large numbers of healthy hPSCs for further experiments.

Figure 2. 3D static suspension culture of hPSC directly from Liquid Nitrogen (LN2)

Start the suspension culture by using healthy and high-quality cells directly from LN2. hPSC-hydrogel aggregates successfully to form healthy spheroids after 1 day in culture. The hPSC spheroids continue to expand from day 1 to 6 (Figure 2A). The resulting hPSC spheroids also show hallmark features of healthy and high-quality stem cell spheroids, i.e., shallow craters or pockmarks. Figure 2B shows that hPSC static suspension cultures from liquid nitrogen are positive for Alkaline Phosphatase, indicating successful expansion of healthy stem cell populations.

Figure 3. Immunofluorescence images of hPSC spheroids with key pluripotent stem cell markers

VitroGel STEM ensures the undifferentiated state of stem cell lines during scale-up. As shown in Figure 3, hPSC aggregates in VitroGel STEM hydrogel and retain pluripotency after 7 days, evidenced by the expression of key pluripotent stem cell markers, SSEA4, OCT4, SOX2, and TRA-1-60.

REFERENCES/PUBLICATIONS

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

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