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Supplementary Materials

This PDF file includes:

  • fig. S1. Microfabricated molds for the assay.
  • fig. S2. Schematics of the gradient generator design.
  • fig. S3. Flow rate of the device and the dynamics within the channel.
  • fig. S4. Simulated flow conditions of the assay.
  • fig. S5. Consistency of gradient concentration in channels over time.
  • fig. S6. Estimation of cell counts after influence of flow.
  • fig. S7. Validation of integrated assay for proliferation with MCF-7 cell lines.
  • fig. S8. Optimization of cancer cell line culture parameters.
  • fig. S9. Phase-contrast images of 2-week cultures under different culture conditions.
  • fig. S10. Proportion of CTCs before culture affected the potential of cluster formation.
  • fig. S11. Fluorescence in situ hybridization of cultured cells.
  • fig. S12. Epithelial CTC counts before and after culture.
  • fig. S13. Screening of doxorubicin in microwell assay using clinical human primary cancer cells cultured from blood samples.
  • fig. S14. Characterization of cultures.
  • fig. S15. Representative images of residual WBC populations.
  • fig. S16. Scanning electron microscopy images demonstrating the densely packed array of microwells to maximize surface for capturing CTCs for culture.
  • fig. S17. Representative image of a microwell containing cells contaminated by RBCs due to inadequate RBC lysis.
  • fig. S18. Optimization of culture medium conditions.
  • table S1. Concentration of a single reagent at each serpentine.
  • table S2. Samples for preliminary validation of procedure.
  • table S3. Samples evaluated for drug screening.
  • table S4. Average percentage of viable CD45− cells in clinical samples.
  • table S5. Disease evaluation of three sets of serial samples with at least one positive culture generated.
  • table S6. CTC counts per milliliter as reported by notable CTC enrichment methods (non–culture-based).
  • Legend for movie S1

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Other Supplementary Material for this manuscript includes the following:

  • movie S1 (.mp4 format). Illustration of seeding, growth, and drug flow.

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