Advertisement

The Surfaceome pp 249-260 | Cite as

Extracellular Matrix Molecule-Based Capture of Mesenchymal Stromal Cells Under Flow

  • Teresa Massam-Wu
  • Stuart A. Cain
  • Cay M. KieltyEmail author
Protocol
  • 1.1k Downloads
Part of the Methods in Molecular Biology book series (MIMB, volume 1722)

Abstract

We present a method to capture mesenchymal stromal cells (MSCs) by adhesion to extracellular matrix (ECM) molecules under flow conditions. The technique simulates a physiological system and exploits the natural biological interactions of cells, through integrin receptors, with their ECM. The system offers an insight into how MSCs could be targeted/localized to the site of interest (graft) following intravenous injection.

Key words

Fibronectin Laminin Adhesion Mesenchymal stromal cell 

Notes

Acknowledgment

This work was supported by the UK Engineering and Physical Sciences Research Council (EPSRC). The Bioimaging Facility microscopes used in this study were purchased with grants from the UK Biotechnology and Biological Sciences Research Council (BBSRC) and the Wellcome Trust, and the University of Manchester Strategic Fund. The Beckman Coulter Cyan ADP used in this study was purchased with a grant from Astra Zeneca. Special thanks to Dr. Peter March for his help with the microscopy, Dr. Christoph Ballestrem and Dr. Alex Carisey for their help with the ibidi pump system, and Mr. Mike Jackson for his help with the flow cytometry.

Supplementary material

Video 1

Mesenchymal stromal cells (MSCs) attachment on a 100 nM laminin -521 coated channel. MSCs begin to attach immediately on 100 nM laminin-521 coated channels after initiation of flow. The density of MSC attachment increases with time. The video is 2.5× the speed of real time and are generated from images from the first 6.5 min of a 10 min experiment (AVI 281,965 kb)

Video 2

Mesenchymal stromal cells (MSCs) attachment on 100 nM fibronectin type III repeats 7–14 (FN III 7–14) coated channel. MSCs begin to attach on a 100 nM FN III 7–14 coated channel after 3 min following initiation of flow. The density of MSC attachment increases with time. The video is 2.5× the speed of real time and are generated from images from the first 6.5 min of a 10 min experiment (AVI 275,602 kb)

References

  1. 1.
    Ball SG, Worthington JJ, Canfield AE, Merry CLR, Kielty CM (2014) Mesenchymal stromal cells: inhibiting PDGF receptors or depleting fibronectin induces mesodermal progenitors with endothelial potential. Stem Cells 32(3):694–705CrossRefPubMedPubMedCentralGoogle Scholar
  2. 2.
    Schindelin J, Arganda-Carreras I, Frise E, Kaynig V, Longair M, Pietzsch S et al (2012) Fiji: an open-source platform for biological-image analysis. Nat Methods 9(7):676–682CrossRefPubMedGoogle Scholar
  3. 3.
    Cain SA, Baldock C, Gallagher J, Morgan A, Bax DV, Weiss AS, Shuttleworth CA, Kielty CM (2005) Fibrillin-1 interactions with heparin: implications for microfibril and elastic fiber assembly. J Biol Chem 280(34):30526–30537CrossRefPubMedGoogle Scholar
  4. 4.
    Marson A, Rock MJ, Cain SA, Freeman LJ, Morgan A, Mellody K, Shuttleworth CA et al (2005) Homotypic fibrillin-1 interactions in microfibril assembly. J Biol Chem 280(6):5013–5021CrossRefPubMedGoogle Scholar
  5. 5.
    Dominici M, Le Blanc K, Mueller I, Slaper-Cortenbach I, Marini FC, Krause DS et al (2006) Minimal criteria for defining multipotent mesenchymal stromal cells. The International Society for Cellular Therapy position statement. Cytotherapy 8(4):315–317CrossRefPubMedGoogle Scholar
  6. 6.
    Kroutvar M, Guttenberg Z (2013) Ibidi Pump System Instructions version 1.5.1, revision 3. http://ibidi.com/fileadmin/products/instruments/I_1090X_PumpSystem/IN_1090X_pump_system.pdf. Accessed 09 Jan 2015

Copyright information

© Springer Science+Business Media, LLC 2018

Authors and Affiliations

  • Teresa Massam-Wu
    • 1
  • Stuart A. Cain
    • 1
  • Cay M. Kielty
    • 1
    Email author
  1. 1.Wellcome Trust Centre for Cell-Matrix Research, Faculty of Life Biology, Medicine and HealthUniversity of ManchesterManchesterUK

Personalised recommendations