Article | Published:

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 12511260 (2012) | Download Citation

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.

Access optionsAccess options

Rent or Buy article

Get time limited or full article access on ReadCube.

from$8.99

All prices are NET prices.

References

  1. 1.

    Development of the vertebrate cornea. Int. Rev. Cytol. 63, 263–322 (1979).

  2. 2.

    & Smooth muscle cell: a key cell for plaque vulnerability regulation? Circ. Res. 98, 448–449 (2006).

  3. 3.

    & Dual origin of intimal cells in cardiac-allograft arteriosclerosis. N. Engl. J. Med. 285, 884–887 (1971).

  4. 4.

    , , & Stromal myofibroblasts are drivers of invasive cancer growth. Int. J. Cancer 123, 2229–2238 (2008).

  5. 5.

    , & The stroma reaction myofibroblast: a key player in the control of tumor cell behavior. Int. J. Dev. Biol. 48, 509–517 (2004).

  6. 6.

    & The tumour microenvironment as a target for chemoprevention. Nat. Rev. Cancer 7, 139–147 (2007).

  7. 7.

    , , , & Epicardium-derived cells contribute a novel population to the myocardial wall and the atrioventricular cushions. Circ. Res. 82, 1043–1052 (1998).

  8. 8.

    Marrow stromal cells as stem cells for nonhematopoietic tissues. Science 276, 71–74 (1997).

  9. 9.

    et al. Blood-borne stem cells differentiate into vascular and cardiac lineages during normal development. Stem Cells Dev. 15, 17–28 (2006).

  10. 10.

    , , , & Bone marrow- derived progenitor cells in pulmonary fibrosis. J. Clin. Invest. 113, 243–52 (2004).

  11. 11.

    , , & Little evidence for developmental plasticity of adult hematopoietic stem cells. Science 297, 2256–2259 (2002).

  12. 12.

    , , , & Fate of the mammalian cardiac neural crest. Science 297, 2256–2259 (2002).

  13. 13.

    & Fibroblasts-a diverse population at the center of it all. Int. Rev. Cell Mol. Biol. 276, 161–214 (2009).

  14. 14.

    , , , & Anatomic demarcation by positional variation in fibroblast gene expression programs. PLoS Genet. 2, e119 (2006).

  15. 15.

    & Structure and function of mesothelial cells. Cancer Treat. Res. 134, 1–19 (2007).

  16. 16.

    & Intrinsic cells: mesothelial cells—central players in regulating inflammation and resolution. Perit. Dial. Int. 29, S21–S27 (2009).

  17. 17.

    & Mesothelial progenitor cells and their potential in tissue engineering. Int. J. Biochem. Cell Biol. 36, 621–642 (2004).

  18. 18.

    , & Mesothelial cell transplantation in myocardial infarction. Int. J. Artif. Organs 30, 541–549 (2007).

  19. 19.

    & Future directions in mesothelial transplantation research. Int. J. Artif. Organs 30, 557–561 (2007).

  20. 20.

    B-1 B cell development. J. Immunol. 177, 2749–2754 (2006).

  21. 21.

    & Molecular cloning of mesothelin, a differentiation antigen present on mesothelium, mesotheliomas, and ovarian cancers. Proc. Natl Acad. Sci. USA 93, 136–140 (1996).

  22. 22.

    et al. MiDReG: a method of mining developmentally regulated genes using Boolean implications. Proc. Natl Acad. Sci. USA 107, 5732–5737 (2010).

  23. 23.

    , , , & Differential regulation of the chick dorsal thoracic dermal progenitors from the medial dermomyotome. Development 129, 4763–4772 (2002).

  24. 24.

    et al. β-catenin activation is necessary and sufficient to specify the dorsal dermal fate in the mouse. Dev. Biol. 296, 164–176 (2006).

  25. 25.

    et al. Role of canonical Wnt signaling/ß-catenin via Dermo1 in cranial dermal cell development. Development 137, 3973–3984 (2010).

  26. 26.

    & Clonal analysis of mouse development reveals a polyclonal origin for yolk sac blood islands. Dev. Cell 11, 519–533 (2006).

  27. 27.

    et al. p53 controls radiation-induced gastrointestinal syndrome in mice independent of apoptosis. Science 327, 593–596 (2010).

  28. 28.

    , , , & Fate of the mammalian cardiac neural crest. Development 127, 1607–1616 (2000).

  29. 29.

    , , , & A global double fluorescent Cre reporter mouse. Genesis 45, 593–605 (2007).

  30. 30.

    et al. A novel transgenic technique that allows specific marking of the neural crest cell lineage in mice. Dev. Biol. 212, 191–203 (1999).

  31. 31.

    et al. The transcriptional control of trunk neural crest induction, survival, and delamination. Dev. Cell 8, 179–192 (2005).

  32. 32.

    et al. Cardiovascular malformations with normal smooth muscle differentiation in neural crest-specific type II TGFβ receptor (Tgfbr2) mutant mice. Dev. Biol. 289, 420–429 (2006).

  33. 33.

    , , , & Physiological migration of hematopoietic stem and progenitor cells. Science 294, 1933–1936 (2001).

  34. 34.

    & Pericardial mesoderm generates a population of smooth muscle cells migrating into the heart along with ingrowth of the pericardial organ. Dev. Biol. 174, 221–232 (1996).

  35. 35.

    Does the subepicardial mesenchyme contribute myocardioblasts to the myocardium of the chick embryo heart? A quail-chick chimera study tracing the fate of the epicardial primordium. Anat. Rec. 255, 212–226 (1999).

  36. 36.

    & The epicardium and epicardially derived cells (EPDCs) as cardiac stem cells. Anat. Rec. A Discov. Mol. Cell Evol. Biol. 276, 43–57 (2004).

  37. 37.

    et al. De novo cardiomyocytes from within the activated adult heart after injury. Nature 474, 640–644 (2011).

  38. 38.

    et al. A myocardial lineage derives from Tbx18 epicardial cells. Nature 454, 104–108 (2008).

  39. 39.

    et al. Epicardial progenitors contribute to the cardiomyocyte lineage in the developing heart. Nature 454, 109–113 (2008).

  40. 40.

    et al. Adult cardiac-resident MSC-like stem cells with a proepicardial origin. Cell Stem Cell 9, 527–540 (2011).

  41. 41.

    , , , & Developmental patterns and characteristics of epicardial cell markers Tbx18 and Wt1 in murine embryonic heart. J. Biomed. Sci. 18, 67 (2011).

  42. 42.

    et al. Tbx18 and the fate of epicardial progenitors. Nature 458, E8–E9 (2009).

  43. 43.

    et al. The Wilms’ tumor gene WT1-GFP knock-in mouse reveals the dynamic regulation of WT1 expression in normal and leukemic hematopoiesis. Leukemia 21, 1783–1791 (2007).

  44. 44.

    , , , & The serosal mesothelium is a major source of smooth muscle cells of the gut vasculature. Development 132, 5317–5328 (2005).

  45. 45.

    et al. Mesothelium contributes to vascular smooth muscle and mesenchyme during lung development. Proc. Natl Acad. Sci. USA 105, 16626–16630 (2008).

  46. 46.

    , , , & Boolean implication networks derived from large scale, whole genome microarray datasets. Gen. Biol. 9, R157 (2008).

Download references

Acknowledgements

We thank A. Mosley for assisting with generating the transgenic MslnCLN 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.

Author information

Affiliations

  1. Institute for Stem Cell Biology and Regenerative Medicine, Departments of Pathology and Developmental Biology, 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

Authors

  1. Search for Yuval Rinkevich in:

  2. Search for Taisuke Mori in:

  3. Search for Debashis Sahoo in:

  4. Search for Pin-Xian Xu in:

  5. Search for John R. Bermingham in:

  6. Search for Irving L. Weissman in:

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.

Competing interests

The authors declare no competing financial interests.

Corresponding author

Correspondence to Yuval Rinkevich.

Supplementary information

PDF files

  1. 1.

    Supplementary Information

    Supplementary Information

Videos

  1. 1.

    Supplementary Information

    Supplementary Information

  2. 2.

    Supplementary Information

    Supplementary Information

About this article

Publication history

Received

Accepted

Published

DOI

https://doi.org/10.1038/ncb2610

Further reading

Newsletter Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing