Gibco StemScale PSC Suspension Medium

A new dimension to PSC culture

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.

  • Maximal expansion capability—achieves 5- to 10-fold expansion per passage and 3x the expansion capability compared to other commercially available PSC suspension media systems
  • Simplified workflow—offers easy media changes without the use of microcarriers or cell strainers during culture or passaging
  • Scalable across formats—supports scale-up of PSC cultures across multiple vessel formats: multiwell plates, shake flasks, and bioreactors
  • Consistency across PSC lines—delivers reliable and consistent spheroid formation while maintaining pluripotency and cell viability

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“StemScale is robust and allows us to replicate the results despite the cell line that we use.”

– C. Miranda, University of Lisbon

Superior expansion capability

Gibco StemScale PSC Suspension Medium supports the growth of pluripotent stem cells in suspension through the self-assembly of spheroids and isideal 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).

Scaling expansion using StemScale Medium

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).

StemScale medium helps to maintain cell viability and pluripotency

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.

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Generate more cells in less time

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.

Pluripotent stem cells grown in StemScale PSC Suspension medium

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.

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“StemScale medium improves upon the efficiency of stem cell aggregation formation.”

– S. Rieck, ViaCyte

Simplified workflow

Simplified workflow for pluripotent stem cell culture maintenance using StemScale Medium

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.

Chart showing similar fold PSC expansion in StemScale medium with every day and every other day feeding.
Chart shows high percent pluripotency marker expression with flexible feeding in StemScale medium

Figure 5. Cultures fed every other day perform similarly to cultures fed daily in fold change (A) and pluripotency (B).

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How-to videos: 3 steps to culturing PSCs in suspension

Watch instructional videos to learn how to culture human pluripotent stem cells in suspension using StemScale PSC Suspension Medium.

Step 1: Initiation of PSC spheroid cultures in StemScale medium
Step 2: Maintenance and feeding of PSC spheroids with 50% media change
Step 3: Passaging PSC spheroids in suspension culture
Step 1: Initiation of PSC spheroid cultures in StemScale medium
Step 2: Maintenance and feeding of PSC spheroids with 50% media change
Step 3: Passaging PSC spheroids in suspension culture

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Expansion across multiple cell lines and vessel formats

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.

Scale-up in suspension culture systems using StemScale PSC Suspension Medium vs. scale-up in adherent culture systems

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.

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Neural and cardiac differentiation from PSC spheroids

Neural induction is initiated directly from PSC spheroids cultured in StemScale PSC Suspension Medium

Neural induction of spheroids

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.

StemScale cultured spheroids are initiated into DA neuron differentiation workflow

Dopaminergic neurons are directly initated 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).

Cardiac organoids differentatiated from PSC spheroids

Cardiomycytes differentiated in suspension from PSC spheroids grown in StemScale medium

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).

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Ordering information

StemScale Medium resources

Find technical and educational resources needed to support your PSC Suspension culture using StemScale PSC Suspension Medium.

Other recommended reagents, consumables, and equipment

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