In vitro chemosensitivity testing of multilayered microcultures

P. E. Pizao, B. Winograd, G. J. Peters, A. Leyva, G. Giaccone, H. M. Pinedo

Research output: Contribution to journalArticlepeer-review

4 Scopus citations


A potential limitation of in vitro microtiter cytotoxicity assays as compared to in vivo antitumor studies is that the complex three-dimensional structure of the solid tumor is lost in monolayer cultures in vitro. We investigated whether more in vivo like cell-cell interactions could be easily and reproducibly obtained in an in vitro cytotoxicity assay. HT29 human colon adenocarcinoma cells were seeded in 96-well microtiter plates with 'V'-shaped wells and allowed to form postconfluent multilayered cultures. Cross-sections of microcultures fixed after 2 and 3 weeks following plating revealed approximately 7 and 35 cell layers respectively. Using a tetrazolium assay to assess cytotoxicity the EC50 (drug concentration which gives absorbance readings 50% lower than those of non-treated wells) of multilayered cultures exposed to doxorubicin for 24 h was 12 times higher (p <0.05) than that determined for subconfluent monolayered cells simultaneouslv exposed to the drug. This system offers an alternative to study the chemosensitivity of three-dimensionally organized cells using semiautomated microtiter plate technology.

Original languageEnglish (US)
Pages (from-to)1319-1322
Number of pages4
JournalAnticancer Research
Issue number4
StatePublished - 1992
Externally publishedYes


  • Cell aggregates
  • Cell culture
  • Cytotoxicity
  • Doxorubicin
  • HT29 cells
  • MTT assay

ASJC Scopus subject areas

  • Cancer Research
  • Oncology


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