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 journalArticle

Abstract

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
Volume12
Issue number4
StatePublished - 1992
Externally publishedYes

Fingerprint

Cell Communication
Pharmaceutical Preparations
Doxorubicin
Reading
Colon
Adenocarcinoma
In Vitro Techniques
Technology
Neoplasms

Keywords

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

ASJC Scopus subject areas

  • Cancer Research
  • Oncology

Cite this

Pizao, P. E., Winograd, B., Peters, G. J., Leyva, A., Giaccone, G., & Pinedo, H. M. (1992). In vitro chemosensitivity testing of multilayered microcultures. Anticancer Research, 12(4), 1319-1322.

In vitro chemosensitivity testing of multilayered microcultures. / Pizao, P. E.; Winograd, B.; Peters, G. J.; Leyva, A.; Giaccone, G.; Pinedo, H. M.

In: Anticancer Research, Vol. 12, No. 4, 1992, p. 1319-1322.

Research output: Contribution to journalArticle

Pizao, PE, Winograd, B, Peters, GJ, Leyva, A, Giaccone, G & Pinedo, HM 1992, 'In vitro chemosensitivity testing of multilayered microcultures', Anticancer Research, vol. 12, no. 4, pp. 1319-1322.
Pizao PE, Winograd B, Peters GJ, Leyva A, Giaccone G, Pinedo HM. In vitro chemosensitivity testing of multilayered microcultures. Anticancer Research. 1992;12(4):1319-1322.
Pizao, P. E. ; Winograd, B. ; Peters, G. J. ; Leyva, A. ; Giaccone, G. ; Pinedo, H. M. / In vitro chemosensitivity testing of multilayered microcultures. In: Anticancer Research. 1992 ; Vol. 12, No. 4. pp. 1319-1322.
@article{8dcaba24fc904948bb40e4490cb5134f,
title = "In vitro chemosensitivity testing of multilayered microcultures",
abstract = "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.",
keywords = "Cell aggregates, Cell culture, Cytotoxicity, Doxorubicin, HT29 cells, MTT assay",
author = "Pizao, {P. E.} and B. Winograd and Peters, {G. J.} and A. Leyva and G. Giaccone and Pinedo, {H. M.}",
year = "1992",
language = "English (US)",
volume = "12",
pages = "1319--1322",
journal = "Anticancer Research",
issn = "0250-7005",
publisher = "International Institute of Anticancer Research",
number = "4",

}

TY - JOUR

T1 - In vitro chemosensitivity testing of multilayered microcultures

AU - Pizao, P. E.

AU - Winograd, B.

AU - Peters, G. J.

AU - Leyva, A.

AU - Giaccone, G.

AU - Pinedo, H. M.

PY - 1992

Y1 - 1992

N2 - 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.

AB - 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.

KW - Cell aggregates

KW - Cell culture

KW - Cytotoxicity

KW - Doxorubicin

KW - HT29 cells

KW - MTT assay

UR - http://www.scopus.com/inward/record.url?scp=0026795361&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0026795361&partnerID=8YFLogxK

M3 - Article

C2 - 1503428

AN - SCOPUS:0026795361

VL - 12

SP - 1319

EP - 1322

JO - Anticancer Research

JF - Anticancer Research

SN - 0250-7005

IS - 4

ER -