Comparison of BioCradle™-L and 2D Culture: Differences in Exosome Production

Using BioCradle™-L enables the potential for stable, continuous exosome production over extended periods.

We present data from a study comparing exosome levels in culture supernatants of adipose-derived mesenchymal stem cells cultured with BioCradle™-L versus 6-well plates.

Results

Per-Cell Exosome Production Efficiency

Using glucose consumption as a cell count indicator, BioCradle™-L showed exosome production efficiency of 445.3 pg/mg per cell on days 9-12, while 2D culture achieved 159.8 pg/mg. BioCradle™-L's per-cell exosome production efficiency was approximately 2.8 times higher than 2D culture, suggesting it provides a more suitable culture environment for exosome production.

Supporting Data 1: Exosome Production Volume

With BioCradle™-L, exosome production continued to increase after day 3, reaching 748.1 pg/mL on days 9-12. In contrast, 2D culture peaked at 393.0 pg/mL on days 3-6, then declined to 274.8 pg/mL by day 12. While both methods showed nearly equivalent exosome production in the early culture period (days 3-6), BioCradle™-L produced approximately 2.7 times more exosomes than 2D culture in the later period (days 9-12).

Supporting Data 2: Glucose Consumption

In BioCradle™-L cultures, glucose consumption increased consistently from start to finish (0.8→1.68 mg/mL), indicating that cells proliferated gradually over time. Meanwhile, 2D culture showed rapid glucose consumption increase (0.7→1.8 mg/mL) followed by immediate plateau, suggesting early cell number saturation.

Materials and Methods

Cell Culture Protocol

Human adipose-derived stem cells (PT-5006, Lonza) were expanded in 2D flasks using serum-free medium (ADSC-5, Kojin Bio). Cells were then seeded at 2.8×10⁴ cells/mL density into 6-well plates (2 mL medium per well) or Erlenmeyer flasks (50 mL medium and 100 μg BioCradle™-L per flask). Starting 16 hours after seeding, Erlenmeyer flasks were shaken for 5 minutes per hour at 120 rpm, and medium in both culture systems was completely replaced every 3 days. Cultures were maintained in a humidity-controlled incubator at 37°C, 5% CO₂.

Exosome Measurement

Measured using CD9/CD63 sandwich ELISA kit (Cosmo Bio).

Glucose Measurement

Measured using BF-8M (Oji Scientific Instruments).

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