3-D Embryonic Stem Cell GFP-Based Fluorescence Assays for High-Throughput Drug Screening, Bioprocess Development, and Biosensing Applications

Posted on by

Summary:

Mammalian, live-cell, fluorescent, kinetic assays are common tools for the pharmaceutical industry, for safety evaluation agencies and even for basic science research as these assays are highly suitable for automatic high-throughput screening and they enable the characterization of rapid cellular events. However, fluorescent signals generated in such assays are usually too weak for in situ measurements. Researchers at The Ohio State University have developed a three-dimensional (3-D) cell culture system that out-performs conventional 2-D cultures which lack proper in vivo tissue functions and are inherently prone to error. The higher cell density in 3-D cell culture shows differences in drug cytotoxicity responses as compared with those from the 2-D cell culture assays and also gives a potential for higher signal to noise ratio (SNR). OSU has designed, built and tested a high throughput, real time, bioactivity assay based on the 3-D culture of GFP-expressing embryonic stem (ES) cells, which can increase SNR at least one order of magnitude and minimize errors due to biological system changes caused by cell activities as compared to conventional 2-D cultures. The 3-D ES cell culture system using inexpensive materials can afford parallel, automated, accurate and long-term (more than one week) cell bioactivity assays. This system has the potential for use as high-throughput biosensors, micro-bioreactor arrays for bioprocess optimization, cytotoxicity assays for drug screening and discovery, and media optimization for expanding and differentiating embryonic stem cells.

Potential Applications:

Markets of interest include the pharmaceutical industry, safety evaluation agencies and basic science research. Applications include high-throughput biosensors, micro-bioreactor arrays for bioprocess development, cytotoxicity assays for drug screening and discovery, and process and media optimization for embryonic stem cell expansion and differentiation.

Advantages:

  • Fluorescent signals are stronger thereby allowing for in situ measurements
  • Improved differenciation in drug cytotoxicity responses
  • Higher signal to noise ratio (SNR) by at least 1 order of magnitude
  • Less errors due to biological system changes
  • Inexpensive materials
  • Can perform parallel, automated, accurate and long-term (>1 week) cell bioactivity assays