Formation of stable small cell number three-dimensional ovarian cancer spheroids using hanging drop arrays for preclinical drug sensitivity assays

Shreya Raghavan, Maria R. Ward, Katelyn R. Rowley, Rachel M. Wold, Shuichi Takayama, Ronald J. Buckanovich, Geeta Mehta

Research output: Contribution to journalArticle

58 Scopus citations

Abstract

Background Ovarian cancer grows and metastasizes from multicellular spheroidal aggregates within the ascites fluid. Multicellular tumor spheroids are therefore physiologically significant 3D in vitro models for ovarian cancer research. Conventional hanging drop cultures require high starting cell numbers, and are tedious for long-term maintenance. In this study, we generate stable, uniform multicellular spheroids using very small number of ovarian cancer cells in a novel 384 well hanging drop array platform. Methods We used novel tumor spheroid platform and two ovarian cancer cell lines (A2780 and OVCAR3) to demonstrate the stable incorporation of as few as 10 cells into a single spheroid. Results Spheroids had uniform geometry, with projected areas (42.60 × 103 μm-475.22 × 103 μm2 for A2780 spheroids and 37.24 × 103 μm2-281.01 × 103 μm2 for OVCAR3 spheroids) that varied as a function of the initial cell seeding density. Phalloidin and nuclear stains indicated cells formed tightly packed spheroids with demarcated boundaries and cell-cell interaction within spheroids. Cells within spheroids demonstrated over 85% viability. 3D tumor spheroids demonstrated greater resistance (70-80% viability) to cisplatin chemotherapy compared to 2D cultures (30-50% viability). Conclusions Ovarian cancer spheroids can be generated from limited cell numbers in high throughput 384 well plates with high viability. Spheroids demonstrate therapeutic resistance relative to cells in traditional 2D culture. Stable incorporation of low cell numbers is advantageous when translating this research to rare patient-derived cells. This system can be used to understand ovarian cancer spheroid biology, as well as carry out preclinical drug sensitivity assays.

Original languageEnglish (US)
Pages (from-to)181-189
Number of pages9
JournalGynecologic oncology
Volume138
Issue number1
DOIs
StatePublished - Jul 1 2015

Keywords

  • 3D culture
  • High throughput
  • Multicellular tumor spheroids
  • Ovarian cancer
  • Preclinical drug testing

ASJC Scopus subject areas

  • Oncology
  • Obstetrics and Gynecology

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