A novel in vitro assay of tumor-initiating cells in xenograft prostate tumors

Christopher R. Silvers, Karin Williams, Linda Salamone, Jiaoti Huang, Craig T. Jordan, Haijun Zhou, Ganesh S. Palapattu

Research output: Contribution to journalArticlepeer-review

3 Scopus citations


BACKGROUND The field of prostate cancer has been stymied by the difficulty of cultivating patient-derived samples in the laboratory. In order to help circumvent this challenge, we sought to develop an in vitro assay of human prostate cancer initiation employing a prostate-associated mesenchymal feeder layer. METHODS Rat seminal vesicle mesenchyme (rSVM) harvested from male neonatal rats was plated in 12-well plates and then irradiated with 30 Gy after ∼75% confluence. Single-cell suspensions of two human non-adherent prostate cancer xenograft lines (TRPC and LAPC9) were then plated on irradiated rSVM. At 3-4 weeks, three-dimensional solid structures, termed glandoids, were harvested and analyzed or transplanted singly into the renal capsule of immunodeficient mice. Animals were assessed for tumor formation 8-12 weeks after engraftment. Finally, clonality assays were performed to determine whether glandoids usually arise from a single cell and are therefore clonal in origin. RESULTS Glandoids form with reliable frequency (1/∼300 plated cells), are constituted by relevant cell types (CK8+, CK5-, PSA+) and after implantation into immunocompromised mice, give rise to tumors that recapitulate original xenograft histology and cell composition; defining a glandoid as a tumor-initiating unit. In addition, assessment of red fluorescent protein (RFP)-labeled glandoids revealed either all red or non-red structures, with few areas of fusion, suggesting glandoids are clonal in origin. CONCLUSIONS The above assay describes an adjunct technique to readily cultivate cells from prostate cancer xenografts in vitro and as such provides a platform on which tumor-initiating cell studies and high-throughput drug discovery may be performed. Prostate 70: 1379-1387, 2010.

Original languageEnglish (US)
Pages (from-to)1379-1387
Number of pages9
Issue number13
StatePublished - Sep 15 2010


  • Cancer stem cell
  • Prostate cancer
  • Seminal vesicle mesenchyme
  • Tumor-initiating cell
  • Xenograft

ASJC Scopus subject areas

  • Urology
  • Oncology


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