ABSTRACT
High Throughput Screening (HTS) with 3D cell models is possible thanks to the recent progress and development in 3D cell culture technologies. Results from multiple studies have demonstrated different drug responses between 2D and 3D cell culture. It is now widely accepted that 3D cell models more accurately represent the physiologic conditions of tumors over 2D cell models. However, there is still a need for more accurate tests that are scalable and better imitate the complex conditions in living tissues. Here, we describe ultrahigh throughput 3D methods of drug response profiling in patient derived primary tumors including melanoma as well as renal cell carcinoma that were tested against the NCI oncologic set of FDA approved drugs. We also tested their autologous patient derived cancer associated fibroblasts, varied the in-vitro conditions using matrix vs matrix free methods and completed this in both 3D vs 2D rendered cancer cells. The result indicates a heterologous response to the drugs based on their genetic background, but not on their maintenance condition. Here, we present the methods and supporting results of the HTS efforts using these 3D of organoids derived from patients. This demonstrated the possibility of using patient derived 3D cells for HTS and expands on our screening capabilities for testing other types of cancer using clinically approved anti-cancer agents to find drugs for potential off label use.
Subject(s)
Antineoplastic Agents , Carcinoma, Renal Cell , Drug Screening Assays, Antitumor , High-Throughput Screening Assays , Melanoma , Humans , Carcinoma, Renal Cell/drug therapy , Carcinoma, Renal Cell/pathology , Carcinoma, Renal Cell/genetics , High-Throughput Screening Assays/methods , Melanoma/drug therapy , Melanoma/pathology , Drug Screening Assays, Antitumor/methods , Antineoplastic Agents/pharmacology , Kidney Neoplasms/drug therapy , Kidney Neoplasms/pathology , Cell Culture Techniques/methods , Cell Line, Tumor , Cell Culture Techniques, Three Dimensional/methods , Drug Evaluation, Preclinical/methodsSubject(s)
COVID-19 , Internship and Residency , Telemedicine , Ambulatory Care Facilities , Humans , PandemicsABSTRACT
The chemical synthesis of gold nanoparticles (NP) by using gold (III) chloride trihydrate (HAuClâ3H2O) and sodium citrate as a reducing agent in aqueous conditions at 100 °C is presented here. Gold nanoparticles areformed by a galvanic replacement mechanism as described by Lee and Messiel. Morphology of gold-NP was analyzed by way of high-resolution transmission electron microscopy; results indicate a six-fold icosahedral symmetry with an average size distribution of 22 nm. In order to understand the mechanical behaviors, like hardness and elastic moduli, gold-NP were subjected to nanoindentation measurements-obtaining a hardness value of 1.72 GPa and elastic modulus of 100 GPa in a 3-5 nm of displacement at the nanoparticle's surface.
