MP31-01 DEVELOPMENT OF NOVEL METASTATIC PROSTATE CANCER CELL LINES BY “ORGANOID” IN VITRO CULTURE TECHNOLOGY

Abstract

You have accessJournal of UrologyProstate Cancer: Basic Research II1 Apr 2014MP31-01 DEVELOPMENT OF NOVEL METASTATIC PROSTATE CANCER CELL LINES BY “ORGANOID” IN VITRO CULTURE TECHNOLOGY Ian Vela, Dong Gao, Anuradha Gopalan, Andrea Sboner, Eva Undvall, Jackline Wanjala, Phillip Iaquinta, John Wongvipat, Catherine Dowling, Wouter Karthaus, Hans Clevers, Stephen B. Solomon, Himisha Beltran, Myriam Kossai, Juan Miguel Mosquera, Mark Rubin, Brett Carver, Howard I. Scher, Charles Sawyers, and Yu Chen Ian VelaIan Vela More articles by this author , Dong GaoDong Gao More articles by this author , Anuradha GopalanAnuradha Gopalan More articles by this author , Andrea SbonerAndrea Sboner More articles by this author , Eva UndvallEva Undvall More articles by this author , Jackline WanjalaJackline Wanjala More articles by this author , Phillip IaquintaPhillip Iaquinta More articles by this author , John WongvipatJohn Wongvipat More articles by this author , Catherine DowlingCatherine Dowling More articles by this author , Wouter KarthausWouter Karthaus More articles by this author , Hans CleversHans Clevers More articles by this author , Stephen B. SolomonStephen B. Solomon More articles by this author , Himisha BeltranHimisha Beltran More articles by this author , Myriam KossaiMyriam Kossai More articles by this author , Juan Miguel MosqueraJuan Miguel Mosquera More articles by this author , Mark RubinMark Rubin More articles by this author , Brett CarverBrett Carver More articles by this author , Howard I. ScherHoward I. Scher More articles by this author , Charles SawyersCharles Sawyers More articles by this author , and Yu ChenYu Chen More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2014.02.910AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES The inability to propagate patient-derived prostate cancer cells in vitro is a major impediment in the mechanistic understanding of tumorigenesis and therapeutic response. In order to generate accurate in vitro models that represent the diversity of in situ prostate cancer, we have developed a three-dimensional “organoid” system to culture metastasis samples and integrated it into our precision medicine workflow of attaining and characterizing pre-treatment biopsies. METHODS Biopsy samples of prostate cancer metastases, both soft tissue and bone, acquired at the time of therapeutic or diagnostic interventions following informed consent and IRB approval were obtained from two institutions. Samples were digested in Type II Collagenase (Gibco) and re-suspended in growth factor reduced Matrigel (BD), plated on plastic, and overlaid with prostate culture media (PCM). PCM consists of serum free Advanced DMEM/F12 (Gibco) with multiple growth factors optimized to propagate benign primary prostate cells. Cultures were maintained at 37°C in 5% CO2. RESULTS In the initial 51 samples, 15 continuous organoid cultures (29%) were established from distinct sites (9/32 bone, 6/19 soft). Tumor content of the biopsy represents a major determinant of organoid growth. Once established, organoids propagate indefinitely with different kinetics (∼48 hr to 1 week doubling time), and can be cryopreserved. Histological analysis shows that the organoids recapitulate the structure of the in situ cancer and genomic analysis using array CGH and whole-exome sequencing (WES) shows the presence of typical copy number alterations including TMPRSS2-ERG interstitial deletion, PTEN loss, CHD1 loss, and AR amplification. WES of two organoid/metastasis pairs shows that the growth conditions do not generate additional mutations. CONCLUSIONS This novel tissue culture technique enables the development of new cell lines derived from metastatic deposits. This advance will facilitate research by availing new and varied cell lines, which will hopefully be more closely aligned to the spectrum of behavior of the clinical disease in comparison to the limited and problematic cell line models currently available. © 2014FiguresReferencesRelatedDetails Volume 191Issue 4SApril 2014Page: e323 Advertisement Copyright & Permissions© 2014MetricsAuthor Information Ian Vela More articles by this author Dong Gao More articles by this author Anuradha Gopalan More articles by this author Andrea Sboner More articles by this author Eva Undvall More articles by this author Jackline Wanjala More articles by this author Phillip Iaquinta More articles by this author John Wongvipat More articles by this author Catherine Dowling More articles by this author Wouter Karthaus More articles by this author Hans Clevers More articles by this author Stephen B. Solomon More articles by this author Himisha Beltran More articles by this author Myriam Kossai More articles by this author Juan Miguel Mosquera More articles by this author Mark Rubin More articles by this author Brett Carver More articles by this author Howard I. Scher More articles by this author Charles Sawyers More articles by this author Yu Chen More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ...