Stem Cell Renewal and Cell-Cell Communication pp 183-193

Part of the Methods in Molecular Biology book series (MIMB, volume 1212) | Cite as

Application of Fluid Mechanical Force to Embryonic Sources of Hemogenic Endothelium and Hematopoietic Stem Cells

Protocol

Abstract

During embryonic development, hemodynamic forces caused by blood flow support vascular remodeling, arterialization of luminal endothelium, and hematopoietic stem cell (HSC) emergence. Previously, we reported that fluid shear stress plays a key role in stimulating nitric oxide (NO) signaling in the aorta-gonad-mesonephros (AGM) and is essential for definitive hematopoiesis. We employed a Dynamic Flow System modified from a cone-and-plate assembly to precisely regulate in vitro exposure of AGM cells to a defined pattern of laminar shear stress. Here, we present the design of a microfluidic platform accessible to any research group that requires small cell numbers and allows for recirculation of paracrine signaling factors with minimal damage to nonadherent hematopoietic progenitors and stem cells. We detail the assembly of the microfluidic platform using commercially available components and provide specific guidance in the use of an emerging standard in the measurement of embryonic HSC potential, intravenous neonatal transplantation.

Keywords:

Hemogenic endothelium Hematopoietic stem cells HSC Biomechanical forces Shear stress AGM 

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Copyright information

© Springer Science+Business Media New York 2014

Authors and Affiliations

  • Nan Li
    • 1
    • 2
  • Miguel F. Diaz
    • 1
    • 2
  • Pamela L. Wenzel
    • 1
    • 2
    • 3
  1. 1.Children’s Regenerative Medicine Program, Department of Pediatric SurgeryUniversity of Texas Medical School at HoustonHoustonUSA
  2. 2.Center for Stem Cell and Regenerative Medicine, The Brown Foundation Institute of Molecular MedicineUniversity of Texas Health Science Center at HoustonHoustonUSA
  3. 3.Department of Pediatric Surgery, Center for Stem Cell and Regenerative Medicine, Institute of Molecular MedicineUniversity of Texas Medical School, SRB-637AHoustonUSA

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