Unit

UNIT 9.25 Advanced Application of CFSE for Cellular Tracking

  1. Jacek M. Witkowski

Published Online: 1 APR 2008

DOI: 10.1002/0471142956.cy0925s44

Current Protocols in Cytometry

Current Protocols in Cytometry

How to Cite

Witkowski, J. M. 2008. Advanced Application of CFSE for Cellular Tracking. Current Protocols in Cytometry. 44:9.25:9.25.1–9.25.8.

Author Information

  1. Medical University of Gdańsk, Gdańsk, Poland

Publication History

  1. Published Online: 1 APR 2008
  2. Published Print: APR 2008

Abstract

This unit proposes a method to extend the already well known dividing-cell-tracking (DCT) cytometric technique based on supravital staining of the lymphocytes with CFSE and allowing them to divide afterwards, beyond simple observation and counting of dividing cells and their generations. Dynamic proliferation parameters that make it possible to determine for in vitro dividing human lymphocytes from various sources, are the actual duration of the pre-division transition period (G0[RIGHTWARDS ARROW]G1), time of a single division, and number of divisions an average dividing cell performs over the time of an experiment, as well as the number of effective precursors giving rise to viable daughter lymphocytes. As the method does not require purification of the lymphocyte population of interest, yet allows the calculations for any cytometrically discernible subpopulation, it presents a powerful tool for detailed analysis of the efficiency of proliferative response of the immune cells. Curr. Protocol. Cytom. 44:9.25.1-9.25.8. © 2008 by John Wiley & Sons, Inc.

Keywords:

  • dividing cell tracking;
  • CFSE;
  • human lymphocytes;
  • cell cycle length;
  • number of divisions;
  • transition time;
  • effective precursors