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

Journal Article (Journal Article)

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 approximately 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/ approximately 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.

Full Text

Duke Authors

Cited Authors

  • Silvers, CR; Williams, K; Salamone, L; Huang, J; Jordan, CT; Zhou, H; Palapattu, GS

Published Date

  • September 15, 2010

Published In

Volume / Issue

  • 70 / 13

Start / End Page

  • 1379 - 1387

PubMed ID

  • 20687210

Pubmed Central ID

  • PMC3808877

Electronic International Standard Serial Number (EISSN)

  • 1097-0045

Digital Object Identifier (DOI)

  • 10.1002/pros.21171

Language

  • eng

Conference Location

  • United States