Growth Inhibition of 3D Tumor Spheres

Image and measure the size of 3D tumor spheroids under multiple drug and growth conditions over time

  1. Directly image tumor spheroids in various microwell formats
  2. Non-invasive bright field imaging allows the user to image the same plate over multiple days
  3. Measure spheroid volume and diameter concurrently with spheroid imaging
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The Celigo imaging cytometer has been developed to fully automate live cell imaging analysis of tumorspheres. This automated morphometric analysis tool significantly reduces the time and effort needed to quantify key aspects of 3D spheres including size, growth, growth tracking over time and response to chemotherapeutics.

automate imaging and analysis of tumorspheres

Drug-Induced Tumor Spheroid Growth Inhibition

Experimental Setup

Experimental Setup Tumor Spheroid Growth Inhibition

4-day old spheroids were treated with compounds for 72 hours. Drug-induced concentration-dependent growth inhibition was obtained.

  • Day 0-4: form spheroids
  • Day 4-7: drug treatment
  • Day 4-14: measure spheroid size

Drug Concentration

drug concentration spheroid growth inhibition

Green calcein AM stained target cells

U 87-MG (giloblastoma) + 17-AGG

U 87-MG (giloblastoma) + 17-AGG

Green calcein AM stained target cells

Live Cell Analysis of Spheroid Growth Inhibition under Drug Treatment and Hypoxia

Experimental Protocol:

  1. Seed 2500 NCI-H460 cells/well in a ULA 96-well plates
  2. On day 4 treat with drug X or vehicle control
  3. Place one plate in hypoxic conditions and one plate at a normoxic condition
  4. On day 4 post-treatment image entire 96-well plate in ~5 minutes

At high drug dose, NCI-H460 3D tumor spheroids showed growth inhibition under hypoxic conditions

NCI-H460 Spheroid at Normoxia, treated with Drug X at 10 µM

spheroid at normoxia drug treated

NCI-H460 cells treated with Drug X and grown in Normoxia vs Hypoxia conditions

NCI-H460 cells treated with Drug X

NCI-H460 Spheroid at Hypoxia, treated with Drug X at 10 µM

spheroid at hypoxia drug treated

Captured bright field images of NCI-H460 spheroids treated with a 10 µM drug and grown at different oxygen conditions show that spheroid size is affected by growth conditions.

By imaging the same plate over multiple days, drug-treated spheroids at hypoxic conditions had smaller spheroid diameters compared to the spheroids grown at normal conditions.