Departmental Papers (BE)

Document Type

Journal Article

Date of this Version

1-2012

Publication Source

Journal of Molecular and Cellular Cardiology

Volume

52

Issue

1

Start Page

105

Last Page

112

DOI

10.1016/j.yjmcc.2011.10.009

Abstract

Clinical trials infusing Bone Marrow Cells (BMCs) into injured hearts have produced measureable improvements in cardiac performance, but were insufficient to improve patient outcomes. Low engraftment rates are cited as probable contributor to limited improvements. To understand the mechanisms that control myocardial engraftment of BMCs following ischemia-reperfusion injury, in isolated–perfused mouse hearts, stop-flow ischemia was followed by variable-duration reperfusion (0–60 min) before addition of labeled syngenic BMCs to the perfusate. After a buffer-only wash, the heart was disaggregated. Retained BMCs (digest) and infused BMCs (aliquot) were compared by flow cytometry for c-kit and CD45 expression to determine the proportion of cell subtypes engrafted versus delivered (selectivity ratio). In these studies, a time-dependent selective retention of c-kit+ cells was apparent starting at 30 min of reperfusion, at which time c-kit+/CD45+ BMCs showed a selectivity ratio of 18 ± 2 (versus 2 ± 1 in sham-ischemic controls). To study the underlying mechanism for this selective retention, neutralizing antibodies for P-selectin or L-selectin were infused into the heart preparation and incubated with BMCs prior to BMC infusion. Blocking P-selectin in ischemic hearts ablated selectivity for c-kit+/CD45+ BMCs at 30 min reperfusion (selectivity ratio of 3 ± 1) while selectivity persisted in the presence of L-selectin neutralization (selectivity ratio of 17 ± 2). To corroborate this finding, a parallel plate flow chamber was used to study capture and rolling dynamics of purified c-kit+ versus c-kit- BMCs on various selectin molecules. C-kit+ BMCs interacted weakly with L-selectin substrates (0.03 ± 0.01% adhered) but adhered strongly to P-selectin (0.28 ± 0.04% adhered). C-kit- BMCs showed intermediate binding regardless of substrate (0.18 ± 0.04% adhered on L-selectin versus 0.17 ± 0.04% adhered on P-selectin). Myocardial ischemia–reperfusion stress induces selective engraftment of c-kit+ bone marrow progenitor cells via P-selectin activation.

Copyright/Permission Statement

© 2012. This manuscript version is made available under the CC-BY-NC-ND 4.0 license http://creativecommons.org/licenses/by-nc-nd/4.0/

Keywords

Ischemia–reperfusion injury, Stem cell therapy, Myocardial infarction, Cell engraftment, CD117, c-kit, CD45, P-selectin, L-selectin

Additional Files

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fig2.pdf (168 kB)
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fig3.pdf (163 kB)
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fig4.pdf (256 kB)
Figure 4

fig5.pdf (1400 kB)
Figure 5

fig6.pdf (118 kB)

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Date Posted: 05 December 2016

This document has been peer reviewed.