The Gibco StemScale™ PSC Suspension Medium is a scalable, easy-to-use medium that supports large-scale generation of human pluripotent stem cells (PSCs) in suspension culture and formulated to address the current technical challenges in aggregating PSC spheroid culture systems.
Dr. Cláudia Miranda shares how StemScale PSC Suspension Medium is enabling her research as a scalable solution for high-fold expansion of human pluripotent stem cells and downstream neural differentiation from PSC spheroids in suspension culture.
“StemScale is robust and allows us to replicate the results despite the cell line that we use.”
Rebecca Sereda talks about her cardiomyocyte research at Harvard University and using StemScale PSC Suspension Medium to create human PSC cardiac organoids at high efficiency in a 3D model system.
“StemScale has provided us a simple and easy way to produce the cells we need to do the science we want.”
Gibco StemScale PSC Suspension Medium supports the growth of pluripotent stem cells in suspension through the self-assembly of spheroids and is ideal for applications where large numbers of high-quality PSCs are required.
StemScale medium delivers 5- to 10-fold expansion per passage and 3x the expansion capability of other media (Figure 1).
Figure 1. Scaling expansion using StemScale medium. After 30 passages and 20 weeks in culture, StemScale medium delivers greater than 10x cell expansion per passage (left plot). When compared against a medium from supplier medium “M”, StemScale medium generates up to 3x expansion across the same period (right plot).
Figure 2. StemScale medium helps to maintain cell viability and pluripotency. StemScale medium delivers superior cell viability after 3 passages as compared to another suspension culture system and maintains high pluripotency after 30 passages.
In comparison to adherent culture, growing PSCs as self-aggregating spheroids in suspension using StemScale Medium allows users to culture greater numbers of cells in a shorter amount of time (Table 1) while reducing the amounts of media and plasticware consumed.
Here we show differences in time and cell number between cells grown in adherent culture and suspension culture starting from a single well of a 6-well plate.
|Suspension scale-up||Adherent scale-up|
|Total days in culture||Culture vessel format||Average cell yield*||Medium consumed||Culture vessel format||Average cell yield*||Medium consumed|
|3 days||6-well plate (single well)||1.5 x 106 cells||6 mL||6-well plate (single well)||1.5 x 106 cells||6 mL|
|7 days||6-well plate (full plate)||18 x 106 cells||30 mL||100 mm dish||9 x 106 cells||48 mL|
|11 days||100 mL bioreactor||150 x 106 cells||200 mL||150 mm dish||22 x 106 cells||72 mL|
|15 days||500 mL bioreactor||750 x 106 cells||1,000 mL||Nunc TripleFlask Treated Cell Culture Flask||78 x 106 cells||800 mL|
|19 days||3 L bioreactor||4.5 x 109 cells||6,000 mL||2-layer Nunc Cell Factory System||300 x 106 cells||1,200 mL|
|23 days||–||–||–||10-layer Nunc Cell Factory System||1.5 x 109 cells||6,000 mL|
|27 days||–||–||–||30-layer Nunc Cell Factory System||4.5 x 109 cells||18,000 mL|
|* Values are estimated from data based on 4 days of expansion per culture vessel format. Values for the 3 L bioreactor and 30-layer Thermo Scientific Nunc Cell Factory System have been extrapolated from the smaller bioreactor and Nunc Cell Factory System formats listed in the table.|
Table 1. Scale-up for suspension vs adherent culture systems. Scale-up in suspension culture systems using StemScale PSC Suspension Medium vs. scale-up in adherent culture systems.
Figure 3. Pluripotent stem cells grown in StemScale medium. Pluripotent stem cells seeded in StemScale PSC Suspension medium maintain optimal spheroid size and viability over 5 days.
“StemScale medium improves upon the efficiency of stem cell aggregation formation.”
– S. Rieck, ViaCyte
Figure 4. Simplified workflow (schematic). Easily adopt adherent cultures to suspension cultures using StemScale medium. StemScale medium enables users to skip feeding days, if desired. After initiation of cultures in StemScale medium, Gibco Episomal iPSCs are fed periodically using a 50% medium exchange, every day or every other day.
Figure 5. Cultures fed every other day perform similarly to cultures fed daily in fold change (A) and pluripotency (B).
Watch instructional videos to learn how to culture human pluripotent stem cells in suspension using StemScale PSC Suspension Medium.
Across four different PSC lines cultured in StemScale Medium, 6- to 11-fold expansion was achieved per passage and 18- to 28-fold expansion was achieved over 15 days.
Figure 6. Suspension vs adherent culture systems. Scale-up in suspension culture systems using StemScale PSC Suspension Medium vs. scale-up in adherent culture systems.
Figure 7. Neural induction of spheroids. Using the PSC Neural Induction Medium (Cat. No. A1647801), large numbers of maturing neural stem cells are initiated directly from PSC spheroids cultured in StemScale medium.
Figure 8. Dopaminergic neurons are directly initiated from PSC spheroids cultured in StemScale medium. In just 25 days, using the PSC Dopaminergic Neuron (DA) Differentiation kit (Cat. No. A3147701) DA neurons differentiated in StemScale medium co-expressed the same amount of FOXA2/Otx2 in suspension culture when compared to a 35-day protocol in adherent culture (data not shown).
Figure 9. Cardiomyocytes differentiated in suspension from PSC spheroids grown in StemScale medium. (A) Undifferentiated hPSC spheres growing in StemScale medium prior to cardiac induction. (B) Spheroids in rotation culture 12 days after start of cardiac induction. (C) Cardiac organoids stained for the cardiac markers Alpha Sarcomeric Actinin (green) and Cardiac Troponin T (red). Nuclei are counterstained with DAPI (blue).
Find technical and educational resources needed to support your PSC Suspension culture using StemScale PSC Suspension Medium.
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