Figure 1. Consistently High Daily Fold Expansion of Human Pluripotent Stem Cells (hPSCs) Is Achieved with the PBS-MINI 0.1 and 0.5 MAG Single-Use Vessels

Daily fold expansions of a human embryonic stem (ES) cell line (H9) and human induced pluripotent stem (iPS) cell line (WLS-1C), cultured in PBS-MINI (A) 0.1 and (B) 0.5 MAG Single-Use Vessels, show consistently high growth. The cells were seeded at a density of 8 x 10⁴ viable cells/mL in 100 mL (for PBS-MINI 0.1 MAG) and 3 - 8 x 10⁴ viable cells/mL in 500 mL (for PBS-MINI 0.5 MAG), and cultured for 4 days using mTeSR™3D (Catalog #03950). Fold expansion of each cell line was comparable to their growth in Nalgene™ filter bottles (Control), when seeded at a density of 1 x 10⁵ viable cells/mL in 30 mL. Error bars are +/- SD (n = 3 for PBS-MINI 0.1 and 0.5 conditions, and n = 2 for control).

Figure 2. Culturing hPSCs in the PBS-MINI 0.1 and 0.5 MAG Single-Use Vessels Achieves High Cell Viability

End-of-passage viabilities of a human ES cell line (H9) and human iPS cell line (WLS-1C) cultured in mTeSR™3D show that the PBS-MINI 0.1 and 0.5 MAG Single-Use Vessels support expansion of hPSCs at (A) 100 mL (with PBS-MINI 0.1 MAG) and (B) 500 mL (with PBS-MINI 0.5 MAG) culture volumes. Error bars are +/- SD (n = 3 for PBS-MINI 0.1 and 0.5 conditions, and n = 2 for control).

Figure 3. Markers of Undifferentiated hPSCs Harvested from the PBS-MINI 0.5 MAG Single-Use Vessels

1C and H9 hPSC lines after 9 passages in the PBS-MINI 0.5 MAG Single-Use Vessels using mTeSR™3D show high OCT-4 and TRA-1-60 expression. Error bars are +/- SD (n = 1).

Figure 4. Morphology of hPSC Aggregates Is Optimal When Using the PBS-MINI 0.1 and 0.5 MAG Single-Use Vessels

hPSCs, expanded in suspension using mTeSR™3D in the PBS-MINI 0.1 and 0.5 MAG Single-Use Vessels, exhibit the typical morphological characteristics of hPSC aggregates in suspension culture, including a mostly spherical shape, clear edges (not smooth or shiny), an even color, and a dimpled or pock-marked appearance. Aggregates are typically 350-400 µm in size. Scale bar = 500 µm.

Figure 5. PBS-MINI 0.1 and 0.5 MAG Single-Use Vessels Demonstrate Consistently High Daily Fold Expansion Compared to Common Alternative Culture Vessels

Daily fold expansion of 7 cell lines in suspension culture vessels, tested from 2019 - 2022. Of the 9 vessels tested, those recommended for robust suspension culture scale-up are the Non Tissue Culture-Treated 6-Well Plate, Nalgene™ Filter Storage Bottle, PBS-MINI 0.1, and PBS-MINI 0.5. These vessels have a consistent mean daily fold expansion above 1.2.