Cell and Tissue Research

, Volume 341, Issue 1, pp 147–158 | Cite as

Integrin expression and integrin-mediated adhesion in vitro of human multipotent stromal cells (MSCs) to endothelial cells from various blood vessels

  • Julie A. Semon
  • Lauren H. Nagy
  • Claire B. Llamas
  • H. Alan Tucker
  • Ryang Hwa Lee
  • Darwin J. Prockop
Regular Article

Abstract

Multipotent mesenchymal stromal cells (MSCs) home to damaged tissue by processes partly regulated by integrins. Integrin subunits expressed by MSCs were identified by flow cytometry (FC), immunocytochemistry (IC), and a panel of integrin-binding antibodies. In subconfluent cultures, over 80% of MSCs expressed integrin subunits β1, β2, and α3, 20%–55% expressed α1, α2, α4, α5, α6, and αV, and about 10% expressed β3 when assayed by FC. None of the cells expressed significant levels of 13 other integrins as assayed by FC, but seven of the 13 integrins were detected by IC: β5, α7, α8, α9, α11, αX, and αD. Expression of some integrins changed with MSC confluency: integrins β3, α1, α3, α5, and αV increased, and α6 decreased. Furthermore, α4 was the only integrin to vary among preparations of MSCs from different donors. The results resolved some discrepancies in the literature concerning integrin expression by MSCs. We also investigated the role of specific integrins in MSC adhesion to endothelial cells (ECs) from the pulmonary artery (HPAEC), cardiac-derived microvasculature (HMVEC-C), and umbilical veins (HUVEC). In experiments with blocking antibodies to beta integrins, anti-β5 reduced MSC adhesion to all types of ECs, anti-β1 to both HUVEC and HPAEC, anti-β3 to HUVEC, and anti-β2 to HMVEC-C. With blocking antibodies to alpha integrins, anti-αX reduced adhesion to HPAEC and HMVEC-C, anti-αV to HPAEC, and both anti-α7 and anti-αD to HMVEC-C. Thus, MSCs use diverse integrins to adhere to EC from various blood vessels in vitro.

Keywords

Multipotent mesenchymal stromal cells Adhesion Integrin Trafficking Homing Human 

Supplementary material

441_2010_994_MOESM1_ESM.doc (56 kb)
Table S1Antibodies used in this study (DOC 55 kb)
441_2010_994_MOESM2_ESM.ppt (535 kb)
Fig. S1Adherence of MSCs to matrices. a About 6×105 MSCs were plated per well of a 24-well plate and incubated for 0–60 min. After incubation, wells were gently washed three times with phosphate-buffered saline (PBS) to remove nonadherent cells, and 10 fields per plate were counted at ×10 magnification. b About 1.5×105 MSCs in 100 µl CCM were added to wells of a 96-well plate pre-coated with various extracellular matrix proteins. After being incubated for 1 h at 37°C, wells were gently washed three times with PBS, and remaining adherent MSCs were labeled with a fluorescent DNA-labeling dye. Adherent cells were measured in a fluorimeter. Values are means±SD, n = 3. *P<0.01 compared with bovine serum albumin (BSA) control (PPT 535 kb)

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Copyright information

© Springer-Verlag 2010

Authors and Affiliations

  • Julie A. Semon
    • 1
  • Lauren H. Nagy
    • 2
  • Claire B. Llamas
    • 1
  • H. Alan Tucker
    • 1
  • Ryang Hwa Lee
    • 3
  • Darwin J. Prockop
    • 1
    • 3
  1. 1.Tulane University Center for Gene TherapyNew OrleansUSA
  2. 2.Department of Medical Microbiology and ImmunologyUniversity of California-DavisDavisUSA
  3. 3.Texas A & M Health Science Center Institute for Regenerative Medicine at Scott & WhiteTempleUSA

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