Scalable Culture Strategies for the Expansion of Patient-Derived Cancer Stem Cell Lines

Figure 3

NSCLC and CRC cell culture in four different microcarriers: PPlus 102-L, Pro-F 102-L, Fact 102-L, and CGEN 102-L. Cells were inoculated at 0.2 × 10⁴ cell/cm² and cultured for 6 days under static culture systems using two different culture media: serum-containing medium (SCM) and serum-free medium (SFM). (a) Fold increase in NSCLC (upper panel) and CRC (lower panel) cell concentration at day 6 of culture on microcarriers using both culture media. Total cell concentration was determined by crystal violet nucleic stain assay. (b) Phase-contrast and fluorescence microscopy images of NSCLC and CRC cells cultured on PPlus 102-L microcarriers. Viability analysis of cultures stained with fluorescein diacetate (FDA—live cells, green) and propidium iodide (PI—dead cells, red). Scale bars: 100 μm. (c) Flow cytometry analysis of NSCLC and CRC cell population at inoculum and after 6 days of culture in microcarriers using serum-free medium. The left panel shows the dot blot of ALDEFLUOR™ assay with an inhibitor (DEAB), and the right panel shows the dot blot without an inhibitor. The ALDH⁺ cell population is identified in green.

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