Skip to main content

Abstract 5363: Aggregation rather than monoclonal expansion explains ovarian cancer spheroid formation

Publication ,  Journal Article
Huang, Z; Lowery, WJ; Berchuck, A; Murphy, SK
Published in: Cancer Research
April 15, 2012

Purpose: It is thought that a subpopulation of ovarian cancer cells possess stem cell like properties and are responsible for both the emergence of cancer and for subsequent chemoresistance and recurrence. A common approach used to enrich for cancer stems cells in vitro involves culture in selective media on low attachment plates resulting in non-attached multicellular spheres. Spheroids are present in ascites fluid of women with ovarian cancer, and may contain stem cells responsible for metastasis and recurrence. Our objective was to address whether spheroids are truly monoclonal in origin and arise from expansion of a single common progenitor cell. Procedures: We cultivated 5 ovarian cancer cell lines (DOV13, HEYC2, OVCAR2, OVCAR3, and PEO4) to confluence in monolayer cultures. The cell lines were chosen for their ability to form spheroids as well as variation in expression of the stem cell marker CD133 (OVCAR2, OVCAR3 CD133+, others negative). The cells were trypsinized to form single cell suspensions and plated at densities ranging from 100 to 5000 cells per plate in stem cell-selective media in ultra-low attachment culture dishes. A Zeiss Axio Observer microscope was used for time-lapse photography of the plated cells at 5-minute intervals over a 24-hour period. In addition, single cell suspensions were subjected to flow activated cell sorting to plate one cell in each well of 96-well Costar ultra-low cluster plates to monitor clonal formation of spheroids. Results: Cells were dissociated and plated under stem cell-selective conditions and 288 images of each cell line were recorded over 24 hours. The cells were found to begin to aggregate within two to four hours, regardless of cell numbers plated. By 24 hours post-plating, there was pronounced aggregation in 5/5 (100%) cell lines with formation of tightly compacted spheroid structures in 4/5 (80%) cell lines. Single cells in the 96-well plates showed no evidence of spheroid formation over a 14-day observation period. Conclusion: We observed spheroid formation in both CD133 positive and negative ovarian cancer cell lines. Formation occured due to cellular aggregation rather than clonal expansion from a single progenitor with stem cell properties. Although cellular aggregation may be relevant to ovarian cancer biology, these data suggest that spheroid formation should be viewed with caution if used as a proxy for monoclonal expansion of ovarian cancer stem cells.Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 5363. doi:1538-7445.AM2012-5363

Duke Scholars

Published In

Cancer Research

DOI

EISSN

1538-7445

ISSN

0008-5472

Publication Date

April 15, 2012

Volume

72

Issue

8_Supplement

Start / End Page

5363 / 5363

Publisher

American Association for Cancer Research (AACR)

Related Subject Headings

  • Oncology & Carcinogenesis
  • 3211 Oncology and carcinogenesis
  • 3101 Biochemistry and cell biology
  • 1112 Oncology and Carcinogenesis
 

Citation

APA
Chicago
ICMJE
MLA
NLM
Huang, Z., Lowery, W. J., Berchuck, A., & Murphy, S. K. (2012). Abstract 5363: Aggregation rather than monoclonal expansion explains ovarian cancer spheroid formation. Cancer Research, 72(8_Supplement), 5363–5363. https://doi.org/10.1158/1538-7445.am2012-5363
Huang, Zhiqing, William J. Lowery, Andrew Berchuck, and Susan K. Murphy. “Abstract 5363: Aggregation rather than monoclonal expansion explains ovarian cancer spheroid formation.” Cancer Research 72, no. 8_Supplement (April 15, 2012): 5363–5363. https://doi.org/10.1158/1538-7445.am2012-5363.
Huang Z, Lowery WJ, Berchuck A, Murphy SK. Abstract 5363: Aggregation rather than monoclonal expansion explains ovarian cancer spheroid formation. Cancer Research. 2012 Apr 15;72(8_Supplement):5363–5363.
Huang, Zhiqing, et al. “Abstract 5363: Aggregation rather than monoclonal expansion explains ovarian cancer spheroid formation.” Cancer Research, vol. 72, no. 8_Supplement, American Association for Cancer Research (AACR), Apr. 2012, pp. 5363–5363. Crossref, doi:10.1158/1538-7445.am2012-5363.
Huang Z, Lowery WJ, Berchuck A, Murphy SK. Abstract 5363: Aggregation rather than monoclonal expansion explains ovarian cancer spheroid formation. Cancer Research. American Association for Cancer Research (AACR); 2012 Apr 15;72(8_Supplement):5363–5363.

Published In

Cancer Research

DOI

EISSN

1538-7445

ISSN

0008-5472

Publication Date

April 15, 2012

Volume

72

Issue

8_Supplement

Start / End Page

5363 / 5363

Publisher

American Association for Cancer Research (AACR)

Related Subject Headings

  • Oncology & Carcinogenesis
  • 3211 Oncology and carcinogenesis
  • 3101 Biochemistry and cell biology
  • 1112 Oncology and Carcinogenesis