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Colony Forming Cell (CFC) Assay for Human Hematopoietic Cells
Published on: December 18, 2010
High throughput cell-based assay of hematopoietic progenitor differentiation
D E Greenwalt1, J Szabo, I Manchel
1BioWhittaker, Inc., Walkersville, MD 21793-0127, USA. dale.greenwalt@biowhittaker.com
Journal of Biomolecular Screening
|January 15, 2002
Summary
A new CELISA assay enables rapid, high-throughput screening of hematopoietic stem cell differentiation. This method accelerates drug discovery and toxicity testing for hematopoiesis-related therapeutics.
Area of Science:
- Hematology
- Stem Cell Biology
- Drug Discovery
Background:
- The traditional clonogenic "colony assay" is low-throughput, limiting its use in drug discovery.
- Assessing drug efficacy on hematopoietic stem cell proliferation and differentiation is crucial.
Purpose of the Study:
- To develop a rapid, high-throughput assay for progenitor cell differentiation.
- To enable efficient drug discovery and toxicity screening in hematopoiesis.
Main Methods:
- Developed a Cell-Enzyme-Linked Immunosorbent Assay (CELISA) using lanthanide-conjugated antibodies.
- Employed time-resolved fluorescence spectroscopy for marker quantification.
- Utilized cell culture-compatible filter plates in 96-well plates for streamlined workflow.
Main Results:
- The CELISA assay quantifies myeloid, erythroid, and megakaryocytic lineages objectively.
- Assay results show strong correlation with the traditional colony assay.
- The entire quantitation process takes approximately 1 hour.
Conclusions:
- The CELISA assay provides a rapid and objective method for assessing hematopoietic progenitor cell differentiation.
- This assay significantly enhances throughput for drug discovery and toxicity screening in hematopoiesis.
- Facilitates objective quantification of multiple hematopoietic lineages.
