Expansion Capability and Its Consistency Across Cell Lines

Basic05 supports strong and reproducible expansion of human iPSCs across multiple cell lines in 3D culture. High growth performance is maintained over passages, while pluripotency remains stable.

(A) Method

Using Basic05 and two competitor media (Medium-A and Medium-B), three human iPSC lines were cultured in 3D under orbital shaking in 125 mL flasks for up to four passages, and growth rate and pluripotency were evaluated.

(B) Cumulative fold expansion in 3D culture of each human iPSC line (four passages).

(C) OCT4-positive rate of each human iPSC line (at each passage).

Basic03 refers to StemFit™ Basic03, a 2D culture medium for hPSC expansion.

Viability and Uniform Spheroid

High cell viability and uniform spheroid formation are critical for stable suspension culture and scalable processes.With Basic05, high cell viability and uniform spheroid formation were observed across multiple human iPSC lines, along with maintenance of a normal karyotype. These results demonstrate that Basic05 enables stable and reproducible culture while maintaining cell quality.

(A) High cell viability across multiple human iPSC lines

Cell viability was evaluated using Vi-CELL and NC-200 across Basic05 and competitor media (Medium-A and Medium-B). Basic05 demonstrated high viability at both Day 1 and Day 3 in cells at passage 3.

(B) Uniform spheroid formation

Basic05 enables uniform spheroid formation with fewer dead cells (Passage 3, Day 1).

(C) Normal karyotype analysis of cells cultured in Basic05

Karyotype analysis at Passage 4 confirmed a normal karyotype in all three iPSC lines cultured in Basic05.

Flexible Workflow

Basic05 provides flexibility in culture scheduling and improves operational efficiency.
Comparable expansion was achieved even when weekend feeding was skipped, in addition to daily feeding conditions. This allows flexible culture schedules based on experimental needs and operational constraints.
As a result, weekend work and labor can be reduced while maintaining stable cell growth. This flexibility is supported not only by the performance of Basic05, but also by the excellent thermal stability of PG-011.

(A) Cumulative cell expansion under different medium change frequencies

Scalability and Usability Among Various Culture Reactors

Basic05 is compatible with a wide range of bioreactors, enabling flexible, platform-independent process development and a smooth transition from research to manufacturing scale, while minimizing re-optimization and technology transfer burden.
Using Basic05, a seamless 2D-to-3D culture process was established and scaled up to 1.5 L, achieving consistent 3–7 fold expansion per passage and stable cell growth from approximately 6 x10⁶ to 1.2 × 10⁹ cells (~200-fold expansion), while maintaining uniform aggregates and high pluripotency.

(A) Scalability Test Method

Over four passages (20 days), scale-up was evaluated by starting from 2D culture and stepwise transitioning to a shaker platform, spinner flask, and vertical-wheel reactor.

(B) Aggregate morphology of human iPSC line
(1383D2) at the end of each passage

(C) Cell growth rate across suspension
culture vessels

(D) Expression analysis of the pluripotency marker OCT4

(E) Cell number over time in 3D culture

Differentiation Potential

(A) Directed differentiation to three germ layers

Basic05 supports the maintenance of differentiation potential after 3D suspension culture.After 3D suspension culture using Basic05, iPSCs demonstrated efficient differentiation into all three germ layers, with lineage-specific marker expression confirmed across all three lineages.

(B) Directed differentiation to hematopoietic progenitor cells via PSC aggregates

Basic05 enables the direct transfer of PSC aggregates from expansion to differentiation, supporting a simplified and scalable workflow.PSC aggregates cultured in suspension with Basic05 were directly differentiated into hematopoietic progenitor cells (HPCs).

2D Performance

Basic05 enables a seamless transition from standard 2D culture to 3D suspension culture using a single medium, improving process development efficiency and reducing the need for re-optimization.In 2D culture, Basic05 showed strong growth and maintained pluripotency, with performance comparable to or better than conventional media.

(A) Method

Basic05 supplemented with 100 ng/mL bFGF was used to culture three human iPSC lines (1383D2, 201B7, and 1210B2) in 2D. Cells were passaged four times, and growth rate and pluripotency (OCT4-positive rate) were evaluated (7 days per passage).

(B) Growth performance across three iPSC lines

Cumulative expansion in 2D culture was evaluated for each cell line using Basic05.Basic05 demonstrated growth performance comparable to or better than Basic03 across all cell lines, confirming its strong performance as a 2D culture medium.

(C) OCT4-positive rate after four passages

Basic03 refers to StemFit™ Basic03,
a 2D culture medium for hPSC expansion.