UNIT 1D.6 High-Throughput Screening Assay for Embryoid Body Differentiation of Human Embryonic Stem Cells
Published Online: 1 MAR 2012
Copyright © 2012 by John Wiley & Sons, Inc.
Lab Protocol Title
Current Protocols in Stem Cell Biology
How to Cite
Outten, J. T., Gadue, P., French, D. L. and Diamond, S. L. 2012. High-Throughput Screening Assay for Embryoid Body Differentiation of Human Embryonic Stem Cells. Current Protocols in Stem Cell Biology. 20:1D.6.1–1D.6.13.
- Published Online: 1 MAR 2012
Serum-free human pluripotent stem cell media offer the potential to develop reproducible clinically applicable differentiation strategies and protocols. The vast array of possible growth factor and cytokine combinations for media formulations makes differentiation protocol optimization both labor and cost-intensive. This unit describes a 96-well plate, 4-color flow cytometry–based screening assay to optimize pluripotent stem cell differentiation protocols. We provide conditions both to differentiate human embryonic stem cells (hESCs) into the three primary germ layers, ectoderm, endoderm, and mesoderm, and to utilize flow cytometry to distinguish between them. This assay exhibits low inter-well variability and can be utilized to efficiently screen a variety of media formulations, reducing cost, incubator space, and labor. Protocols can be adapted to a variety of differentiation stages and lineages. Curr. Protoc. Stem Cell Biol. 20:1D.6.1-1D.6.13. © 2012 by John Wiley & Sons, Inc.
- human embryonic stem cells;
- high throughput;
- embryoid body;
- suspension culture