Abstract LB250: Development of organoid raft cultures of cervical, breast and glioblastoma tumors as quick economical ex vivo human cancer models for pre-clinical drug evaluation

Abstract

Abstract Background: In the past few years, 3D organoid cultures of patient-derived tumors or patient-derived xenografts have gained significant attention as faster and more economical ex vivo alternatives to animal models for the pre-clinical evaluation of therapeutics. We reported previously that raft cultures of ex vivo epithelial warts, normal human epithelia from various anatomic sites, as well as cancer cell lines (cervical and melanoma) grown at the liquid:air interface recapitulate parental tissue phenotypes. Here we describe adaptation of the above technique to develop Organoid Raft Culture (ORC) of tumors of various origin and validation as preclinical models for drug evaluation. Methods: A stromal equivalent (SE) consisting of buffered rat-tail collagen and J2 mouse fibroblasts was prepared in 24-well tissue culture plates. Freshly harvested tumors from patients or patient-derived xenografts of cervical cancers were minced to <2 mm pieces, mixed with buffered rat tail collagen mixture, assembled on the SE and cultured submerged for 2 days in simple raft culture media (DMEM, F12, 10% FBS, growth factors). This assembly was then raised on stainless steel grids in a 6-well plate and cultured at liquid:air interface for variable times (10 days to over 2-3 months). Tumors were harvested, formalin-fixed and paraffin-embedded for analyses by histology and immunohistochemistry. Results: The cervical ORCs maintained the heterogeneity of the parental tumor or xenografts. We evaluated cytotoxicity of vorinostat and CFI402257 on ORCs of cervical cancer-derived PDX models by TUNEL assays and immunohistochemistry. The ex-vivo data was validated in experiments with the in vivo PDXs of cervical cancers. We also successfully developed ORCs of mouse xenografts of syngeneic breast tumor cell lines, and organoids from glioblastomas. Conclusions: Organoid Raft Cultures of patient- or mouse-derived tumors and xenografts of cervical, breast and glioblastoma, recapitulate the morphology of parent tumors, can be passaged multiple times, are suitable for bio-banking and expedite drug evaluation. This method does not require costly growth components and multiple cultures could be easily performed. The PDX derived ORC platform would allow rapid evaluation of drug candidates against many types of solid tumors. Acknowledgement: R21CA216789 grant to NSB. Citation Format: Nilam Sanjib Banerjee, Dianne W. Moore, Abhisek Gangrade, Donald J. Buchsbaum, Luise Burt Nabors, Thomas R. Broker, Louise T. Chow. Development of organoid raft cultures of cervical, breast and glioblastoma tumors as quick economical ex vivo human cancer models for pre-clinical drug evaluation [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr LB250.