An Organotypic High Throughput System for Characterization of Drug Sensitivity of Primary Multiple Myeloma Cells

J Vis Exp. 2015 Jul 15;(101):e53070. doi: 10.3791/53070.

Abstract

In this work we describe a novel approach that combines ex vivo drug sensitivity assays and digital image analysis to estimate chemosensitivity and heterogeneity of patient-derived multiple myeloma (MM) cells. This approach consists in seeding primary MM cells freshly extracted from bone marrow aspirates into microfluidic chambers implemented in multi-well plates, each consisting of a reconstruction of the bone marrow microenvironment, including extracellular matrix (collagen or basement membrane matrix) and stroma (patient-derived mesenchymal stem cells) or human-derived endothelial cells (HUVECs). The chambers are drugged with different agents and concentrations, and are imaged sequentially for 96 hr through bright field microscopy, in a motorized microscope equipped with a digital camera. Digital image analysis software detects live and dead cells from presence or absence of membrane motion, and generates curves of change in viability as a function of drug concentration and exposure time. We use a computational model to determine the parameters of chemosensitivity of the tumor population to each drug, as well as the number of sub-populations present as a measure of tumor heterogeneity. These patient-tailored models can then be used to simulate therapeutic regimens and estimate clinical response.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't
  • Video-Audio Media

MeSH terms

  • Antineoplastic Agents / pharmacology*
  • Bone Marrow / pathology
  • Bone Marrow Cells / pathology
  • Cell Line, Tumor
  • Drug Screening Assays, Antitumor / methods*
  • Endothelial Cells / cytology
  • High-Throughput Screening Assays / methods*
  • Humans
  • Lab-On-A-Chip Devices
  • Mesenchymal Stem Cells / cytology
  • Multiple Myeloma / drug therapy*
  • Multiple Myeloma / pathology
  • Tumor Cells, Cultured

Substances

  • Antineoplastic Agents