Isolation of human umbilical cord blood aldehyde dehydrogenase-expressing progenitor cells that modulate vascular regenerative functions in vitro and in vivo

Curr Protoc Stem Cell Biol. 2013:Chapter 2:Unit 2A.10. doi: 10.1002/9780470151808.sc02a10s25.

Abstract

This unit describes the isolation and application of human umbilical cord blood progenitor cells to modulate vascular regenerative functions using in vitro co-culture systems and in vivo transplantation models. Using aldehyde dehydrogenase as a marker of stem cell function, blood-derived progenitors can be efficiently purified form human umbilical cord blood using flow cytometry. We describe in vitro approaches to measure cell-mediated effects on the survival, proliferation, and tube-forming function of endothelial cells using growth-rate assays and Matrigel tube-forming assays. Additionally, we provide a detailed protocol for inducing acute unilateral hindlimb ischemia in immune-deficient mice to assess progenitor cell-modulated effects on vascular regeneration by tracking the recovery of blood flow using noninvasive laser Doppler perfusion imaging. Collectively, we present combined in vitro and in vivo transplantation strategies for the pre-clinical assessment of human progenitor cell-based therapies to treat ischemic disease.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Aldehyde Dehydrogenase / metabolism*
  • Animals
  • Blood Vessels / drug effects
  • Blood Vessels / physiology*
  • Cell Proliferation / drug effects
  • Cell Separation / methods*
  • Cell Survival / drug effects
  • Coculture Techniques
  • Culture Media, Conditioned / pharmacology
  • Femoral Artery / drug effects
  • Fetal Blood / cytology*
  • Flow Cytometry
  • Human Umbilical Vein Endothelial Cells / cytology
  • Human Umbilical Vein Endothelial Cells / drug effects
  • Human Umbilical Vein Endothelial Cells / metabolism
  • Humans
  • Ligation
  • Mice
  • Paracrine Communication / drug effects
  • Regeneration / drug effects
  • Regeneration / physiology*
  • Stem Cell Transplantation
  • Stem Cells / cytology
  • Stem Cells / drug effects
  • Stem Cells / enzymology*

Substances

  • Culture Media, Conditioned
  • Aldehyde Dehydrogenase