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High-resolution mass spectrometric analysis of the secretome from mouse lung endothelial progenitor cells

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Abstract

Recently, we isolated and characterized resident endothelial progenitor cells from the lungs of adult mice. These cells have a high proliferation potential, are not transformed and can differentiate into blood- and lymph-vascular endothelial cells under in vitro and in vivo conditions. Here we studied the secretome of these cells by nanoflow liquid chromatographic mass spectrometry (LC–MS). For analysis, 3-day conditioned serum-free media were used. We found 133 proteins belonging to the categories of membrane-bound or secreted proteins. Thereby, several of the membrane-bound proteins also existed as released variants. Thirty-five proteins from this group are well known as endothelial cell- or angiogenesis-related proteins. The MS analysis of the secretome was supplemented and confirmed by fluorescence activated cell sorting analyses, ELISA measurements and immunocytological studies of selected proteins. The secretome data presented in this study provides a platform for the in-depth analysis of endothelial progenitor cells and characterizes potential cellular markers and signaling components in hem- and lymphangiogeneis.

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Acknowledgments

We thank Dr. Dietmar Vestweber (Muenster) for the ESAM and VE-cadherin antibodies, Dr. Sirpa Jalkanen (Turku) for the Stabilin-1/CLEVER1 antibody, Dr. Alfonso Colombatti (Udine) for the Emilin1 antibody and Dr. Urban Deutsch (Bern) for the KDR/VEGFR-2 and -3 antibodies. Especially we would like to thank Mrs. B. Pawletta for ELISA measurements and cell culture experiments.

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Authors and Affiliations

  1. Department of Gene Regulation, HZI, Build. D, Inhoffenstr. 7, 38124, Braunschweig, Germany

    Katherina Hemmen, Friederike Behler, Hauke Dieken & Herbert A. Weich

  2. Proteome Research Group, Helmholtz Centre for Infection Research, Braunschweig, Germany

    Tobias Reinl & Lothar Jänsch

  3. Department Anatomy and Cell Biology, Georg-August-University, Goettingen, Germany

    Kerstin Buttler & Jörg Wilting

Authors
  1. Katherina Hemmen
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  2. Tobias Reinl
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  3. Kerstin Buttler
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  4. Friederike Behler
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  5. Hauke Dieken
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  6. Lothar Jänsch
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  7. Jörg Wilting
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  8. Herbert A. Weich
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Corresponding author

Correspondence to Herbert A. Weich.

Additional information

Katherina Hemmen and Tobias Reinl contributed equally.

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Hemmen, K., Reinl, T., Buttler, K. et al. High-resolution mass spectrometric analysis of the secretome from mouse lung endothelial progenitor cells. Angiogenesis 14, 163–172 (2011). https://doi.org/10.1007/s10456-011-9200-x

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  • DOI: https://doi.org/10.1007/s10456-011-9200-x

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