Abstract 4672: Development of 2D - cell strain and 3D - tumor spheroid models for Precision Medicine
Authors: Pauli Chantal , Pauwels Jonathan , Kossai Myriam , Steklov Nikolai , Sboner Andrea , Rao Rema , Hennrick Kenneth , Robinson Brian , Mosquera Juan Miguel , Beltran Himisha , Rubin Mark A. 
Published: August 2015

Background

Precision oncology is a clinical approach aimed towards tailoring treatment strategies for patients based on the genetic profile of each patient's malignancy. Available cell line models are greatly limited in recapitulating the genetic profile of the tumor and tumor microenvironment of patients and therefore limit preclinical evaluation of new targeted agents. Furthermore, a high failure rate of drug candidates can be attributed in part to use of two dimensional (2D) monolayer cultures as the initial screening method that frequently produces inaccurate results and does not predict chemoresistance. The ability of three dimensional (3D) cultured tumor cells derived directly from patients to recapitulate tumor-like microenvironment suggests that engineered tumor models may provide more accurate and reproducible information for drug testing. At the Institute for Precision Medicine, we grow patient-derived 2D cell strains and engineer 3D tumor spheroids for patient`s tumor specific drug testing.

Design

Fresh tissue samples from 30 different solid tumors were digested in Type II or IV Collagenase (Gibco) and then re-suspended in growth factor reduced Matrigel (BD), plated on plastic, and overlaid with cell type specific cell growth media tsCM, which consists of either serum free Advanced DMEM/F12 (Gibco) with multiple growth factors optimized to propagate benign primary tumor cells, or FBS enriched DMEM/F12 media. Cultures were maintained at 37°C in 5% CO2. Once established, early cell strain culture passage and tumor spheroids size &lt; 500 μm were cryopreserved. Specific drug testing was performed following 3 models: 2D monolayer culture, 2D monolayer culture converted in a 3D system using the hanging drop system from Biomatrix 3D®, and engineered 3D tumor spheroids.

Results

Our success rate in generating 2D- cell strains and 3D tumor spheroids is about 30%-40%. In the initial 30 samples, we were able to establish 11 continuous 2D and 3D tumor spheroid cultures derived from distinct primary and metastatic cancers including prostate, kidney, bladder and endometrium, as well as sarcomas and a neuroendocrine tumor. Tumor content in biopsy represents a major factor for successful growth. Histology and cytology examination demonstrates that 3D tumor spheroids recapitulate harvested tumors. Next generation sequencing shows the presence of same genomic alterations compared with original tumor without generating additional mutations.

Conclusion

We have developed individual patient-derived 2D tumor cell strains and 3D engineered tumor spheroids as well as models for patient`s tumor specific drug testing. This effort has been implemented for Precision Care and Clinical Trials at our Institution.

Citation Format: Chantal Pauli, Jonathan Pauwels, Myriam Kossai, Nikolai Steklov, Andrea Sboner, Rema Rao, Kenneth Hennrick, Brian Robinson, Juan Miguel Mosquera, Himisha Beltran, Mark A. Rubin. Development of 2D - cell strain and 3D - tumor spheroid models for Precision Medicine. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 4672. doi:10.1158/1538-7445.AM2015-4672