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Establishment and Analysis of Three-Dimensional (3D) Organoids Derived from Patient Prostate Cancer Bone Metastasis Specimens and their Xenografts.
Lee, Sanghee; Burner, Danielle N; Mendoza, Theresa R; Muldong, Michelle T; Arreola, Catalina; Wu, Christina N; Cacalano, Nicholas A; Kulidjian, Anna A; Kane, Christopher J; Jamieson, Christina A M.
Afiliación
  • Lee S; Department of Urology, University of California, San Diego; Moores Cancer Center, University of California, San Diego.
  • Burner DN; Department of Urology, University of California, San Diego; Moores Cancer Center, University of California, San Diego.
  • Mendoza TR; Department of Urology, University of California, San Diego; Moores Cancer Center, University of California, San Diego.
  • Muldong MT; Department of Urology, University of California, San Diego; Moores Cancer Center, University of California, San Diego.
  • Arreola C; Department of Urology, University of California, San Diego; Moores Cancer Center, University of California, San Diego.
  • Wu CN; Moores Cancer Center, University of California, San Diego; Department of Medicine, University of California, San Diego.
  • Cacalano NA; Department of Radiation Oncology, University of California, Los Angeles.
  • Kulidjian AA; Moores Cancer Center, University of California, San Diego; Department of Orthopedic Surgery, University of California, San Diego.
  • Kane CJ; Department of Urology, University of California, San Diego; Moores Cancer Center, University of California, San Diego.
  • Jamieson CAM; Department of Urology, University of California, San Diego; Moores Cancer Center, University of California, San Diego; camjamieson@ucsd.edu.
J Vis Exp ; (156)2020 02 03.
Article en En | MEDLINE | ID: mdl-32065165
Three-dimensional (3D) culture of organoids from tumor specimens of human patients and patient-derived xenograft (PDX) models of prostate cancer, referred to as patient-derived organoids (PDO), are an invaluable resource for studying the mechanism of tumorigenesis and metastasis of prostate cancer. Their main advantage is that they maintain the distinctive genomic and functional heterogeneity of the original tissue compared to conventional cell lines that do not. Furthermore, 3D cultures of PDO can be used to predict the effects of drug treatment on individual patients and are a step towards personalized medicine. Despite these advantages, few groups routinely use this method in part because of the extensive optimization of PDO culture conditions that may be required for different patient samples. We previously demonstrated that our prostate cancer bone metastasis PDX model, PCSD1, recapitulated the resistance of the donor patient's bone metastasis to anti-androgen therapy. We used PCSD1 3D organoids to characterize further the mechanisms of anti-androgen resistance. Following an overview of currently published studies of PDX and PDO models, we describe a step-by-step protocol for 3D culture of PDO using domed or floating basement membrane (e.g., Matrigel) spheres in optimized culture conditions. In vivo stitch imaging and cell processing for histology are also described. This protocol can be further optimized for other applications including western blot, co-culture, etc. and can be used to explore characteristics of 3D cultured PDO pertaining to drug resistance, tumorigenesis, metastasis and therapeutics.
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Neoplasias de la Próstata / Neoplasias Óseas / Organoides / Técnicas de Cultivo de Tejidos Tipo de estudio: Guideline / Prognostic_studies Límite: Humans / Male Idioma: En Revista: J Vis Exp Año: 2020 Tipo del documento: Article Pais de publicación: Estados Unidos
Texto completo
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Neoplasias de la Próstata / Neoplasias Óseas / Organoides / Técnicas de Cultivo de Tejidos Tipo de estudio: Guideline / Prognostic_studies Límite: Humans / Male Idioma: En Revista: J Vis Exp Año: 2020 Tipo del documento: Article Pais de publicación: Estados Unidos