Unit

UNIT 1B.7 Preparation of Defined Human Embryonic Stem Cell Populations for Transcriptional Profiling

  1. Qi Zhou1,
  2. Hun Chy1,
  3. Andrew L. Laslett1,2

Published Online: 1 JUL 2010

DOI: 10.1002/9780470151808.sc01b07s14

Lab Protocol Title

Current Protocols in Stem Cell Biology

Current Protocols in Stem Cell Biology

Additional Information

How to Cite

Zhou, Q., Chy, H. and Laslett, A. L. 2010. Preparation of Defined Human Embryonic Stem Cell Populations for Transcriptional Profiling. Current Protocols in Stem Cell Biology. 14:B:1B.7:1B.7.1–1B.7.15.

Author Information

  1. 1

    Commonwealth Scientific and Industrial Research Organization (CSIRO), Molecular and Health Technologies, Clayton, Australia

  2. 2

    Department of Anatomy and Developmental Biology, Monash University, Clayton, Australia

Publication History

  1. Published Online: 1 JUL 2010
  2. Published Print: NOV 2010
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Abstract

This unit describes a useful approach to preparing highly reproducible samples of human embryonic stem cell (hESC) total RNA suitable for transcriptional profiling from heterogeneous mixtures of cells containing undifferentiated hESC and differentiated cell types. In this unit, fluorescence-activated cell sorting (FACS) is used to sub-fractionate hESC populations on the basis of their levels of co-expression of two previously published hESC surface markers, CD9(TG30) and GCTM-2. This sub-fractionation allows for the separation of undifferentiated hESC (CD9hi, GCTM-2hi) from the early stages in hESC differentiation (CD9neg or low, GCTM-2neg or low). Curr. Protoc. Stem Cell Biol. 14:1B.7.1-1B.7.15. © 2010 by John Wiley & Sons, Inc.

Keywords:

  • human embryonic stem cells;
  • cell surface markers;
  • fluorescence-activated cell sorting
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