3D cell subculturing pillar dish for pharmacogenetic analysis and high-throughput screening

Sang Yun Lee, Hyun Ju Hwang, You Jin Song, Dayoung Lee, Bosung Ku, Jason K. Sa, Dong Woo Lee

Research output: Contribution to journalArticlepeer-review


A pillar dishe for subculture of 3D cultured cells on hydrogel spots (Matrigel and alginate) have been developed. Cells cultured in 3D in an extracellular matrix (ECM) can retain their intrinsic properties, but cells cultured in 2D lose their intrinsic properties as the cells stick to the bottom of the well. Previously, cells and ECM spots were dispensed on a conventional culture dish for 3D cultivation. However, as the spot shape and location depended on user handling, pillars were added to the dish to realize uniform spot shape and stable subculture, supporting 3D cell culture-based high-throughput screening (HTS). Matrigel and alginate were used as ECMs during 6-passage subculture. The growth rate of lung cancer cell (A549) was higher on Matrigel than on alginate. Cancer cell was subcultured in three dimensions in the proposed pillar dish and used for drug screening and differential gene expression analysis. Interestingly, stemness markers, which are unique characteristics of lung cancer cells inducing drug resistance, were upregulated in 3D-subcultured cells compared with those in 2D-subcultured cells. Additionally, the PI3K/Akt/mTOR, VEGFR1/2, and Wnt pathways, which are promising therapeutic targets for lung cancer, were activated, showing high drug sensitivity under 3D-HTS using the 3D-subcultured cells.

Original languageEnglish
Article number100793
JournalMaterials Today Bio
Publication statusPublished - 2023 Dec

Bibliographical note

Publisher Copyright:
© 2023 The Authors


  • 3D cell culture
  • Cell culture dish
  • High-throughput screening
  • Micropillar and well chips

ASJC Scopus subject areas

  • Biotechnology
  • Bioengineering
  • Biomaterials
  • Biomedical Engineering
  • Molecular Biology
  • Cell Biology


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