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Identification and prospective isolation of a mesothelial precursor lineage giving rise to smooth muscle cells and fibroblasts for mammalian internal organs, and their vasculature

Nature Cell Biology volume 14, pages 1251–1260 (2012)Cite this article

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Abstract

Fibroblasts and smooth muscle cells (FSMCs) are principal cell types of connective and adventitial tissues that participate in the development, physiology and pathology of internal organs, with incompletely defined cellular origins. Here, we identify and prospectively isolate from the mesothelium a mouse cell lineage that is committed to FSMCs. The mesothelium is an epithelial monolayer covering the vertebrate thoracic and abdominal cavities and internal organs. Time-lapse imaging and transplantation experiments reveal robust generation of FSMCs from the mesothelium. By targeting mesothelin (MSLN), a surface marker expressed on mesothelial cells, we identify and isolate precursors capable of clonally generating FSMCs. Using a genetic lineage tracing approach, we show that embryonic and adult mesothelium represents a common lineage to trunk FSMCs, and trunk vasculature, with minimal contributions from neural crest, or circulating cells. The isolation of FSMC precursors enables the examination of multiple aspects of smooth muscle and fibroblast biology as well as the prospective isolation of these precursors for potential regenerative medicine purposes.

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Figure 1: Derivation of FSMCs from cultured mesothelium.
Figure 2: Derivation of FSMCs from transplantation of mesothelium in vivo.
Figure 3: Flow cytometry, in vitro clonal analysis and differentiation of MSLN+Lin− cells.
Figure 4: Genetic lineage tracing of fibroblasts within internal organs.
Figure 5: Genetic lineage tracing of smooth muscle within internal organs, and their vasculature.
Figure 6: Polyclonal origins for smooth muscle revealed by clonal analysis of tetrachimaeric mice.
Figure 7: Polyclonal contributions of MSLN+ precursors to smooth muscle during the first postnatal month.
Figure 8: Developmental restriction of MSLN+ precursors to FSMCs following sublethal irradiation.

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Acknowledgements

We thank A. Mosley for assisting with generating the transgenic M s l nCLN mice, and R. Kopito, M. Brandeis and K. Bersuker for usage and their assistance with the time-lapse video. We thank T. Violante for her assistance with sectioning, D. Hunter for his assistance with histochemical staining and G. Paz and D. Montoro for their assistance with figure preparation. The CreERT2IRES–lacZ–PGK–neo cassette was a gift from R. J. Gilbertson (St. Jude Children’s Research Hospital, USA). This work was supported in part by a grant from the California Institute of Regenerative Medicine (RC1 00354 to I.L.W.), the Smith Family Trust (to I.L.W.) and from the National Institutes of Health (RO1 DK064640 to P.X.X.). Y.R. is supported by the Human Frontier Science Program (HFSP) Long Term Fellowship, and the Machiah Foundation Fellowship.

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Authors and Affiliations

  1. Departments of Pathology and Developmental Biology, Institute for Stem Cell Biology and Regenerative Medicine, Stanford University School of Medicine, Stanford, California 94305, USA

    Yuval Rinkevich, Taisuke Mori, Debashis Sahoo & Irving L. Weissman

  2. Division of Molecular Pathology, National Cancer Research Institute, Tokyo 104-0045, Japan

    Taisuke Mori

  3. Department of Genetics and Genomic Sciences, Mount Sinai School of Medicine of NYU, New York, New York 10029, USA

    Pin-Xian Xu

  4. McLaughlin Research Institute, 1520 23rd Street South, Great Falls, Montana 59405, USA

    John R. Bermingham Jr

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Contributions

Y.R. designed and performed the experiments, with suggestions from I.L.W. Y.R. imaged and analysed the data from all experiments. T.M. created the MSLN–CreER construct, with help from Y.R. D.S. contributed the Boolean analysis. P-X.X. contributed the tetrachimaera mouse models. J.R.B. contributed the PoCre transgenic mice model. Y.R. and I.L.W. wrote the manuscript.

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Correspondence to Yuval Rinkevich.

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Rinkevich, Y., Mori, T., Sahoo, D. et al. Identification and prospective isolation of a mesothelial precursor lineage giving rise to smooth muscle cells and fibroblasts for mammalian internal organs, and their vasculature. Nat Cell Biol 14, 1251–1260 (2012). https://doi.org/10.1038/ncb2610

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