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

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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.

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Author notes

  1. Darwin J. Prockop

    Present address: Texas A & M Health Science Center Institute for Regenerative Medicine at Scott & White, 5701 Airport Road, Module C, Temple, TX, 76502, USA

Authors and Affiliations

  1. Tulane University Center for Gene Therapy, 1430 Tulane Avenue, New Orleans, LA, 70112, USA

    Julie A. Semon, Claire B. Llamas, H. Alan Tucker & Darwin J. Prockop

  2. Department of Medical Microbiology and Immunology, University of California-Davis, 1 Shields Avenue, Davis, CA, 95616, USA

    Lauren H. Nagy

  3. Texas A & M Health Science Center Institute for Regenerative Medicine at Scott & White, 5701 Airport Road, Module C, Temple, TX, 76502, USA

    Ryang Hwa Lee

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  1. Julie A. Semon
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  2. Lauren H. Nagy
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Correspondence to Darwin J. Prockop.

Additional information

J.A.S. and D.J.P. designed the research, J.A.S., R.H.L., and D.J.P. analyzed the data, J.A.S and D.J.P wrote the paper, and J.A.S., L.N., C.L., and A.T. performed the experiments. The authors declare that they have no competing financial interests.

This work was supported in part by NIH grants P40 RR 17447 and P01 HL 075161.

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Table S1

Antibodies used in this study (DOC 55 kb)

Fig. S1

Adherence 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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Semon, J.A., Nagy, L.H., Llamas, C.B. et al. Integrin expression and integrin-mediated adhesion in vitro of human multipotent stromal cells (MSCs) to endothelial cells from various blood vessels. Cell Tissue Res 341, 147–158 (2010). https://doi.org/10.1007/s00441-010-0994-4

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