Mesenchymal and haematopoietic stem cells form a unique bone marrow niche

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Abstract

The cellular constituents forming the haematopoietic stem cell (HSC) niche in the bone marrow are unclear, with studies implicating osteoblasts, endothelial and perivascular cells. Here we demonstrate that mesenchymal stem cells (MSCs), identified using nestin expression, constitute an essential HSC niche component. Nestin+ MSCs contain all the bone-marrow colony-forming-unit fibroblastic activity and can be propagated as non-adherent ‘mesenspheres’ that can self-renew and expand in serial transplantations. Nestin+ MSCs are spatially associated with HSCs and adrenergic nerve fibres, and highly express HSC maintenance genes. These genes, and others triggering osteoblastic differentiation, are selectively downregulated during enforced HSC mobilization or β3 adrenoreceptor activation. Whereas parathormone administration doubles the number of bone marrow nestin+ cells and favours their osteoblastic differentiation, in vivo nestin+ cell depletion rapidly reduces HSC content in the bone marrow. Purified HSCs home near nestin+ MSCs in the bone marrow of lethally irradiated mice, whereas in vivo nestin+ cell depletion significantly reduces bone marrow homing of haematopoietic progenitors. These results uncover an unprecedented partnership between two distinct somatic stem-cell types and are indicative of a unique niche in the bone marrow made of heterotypic stem-cell pairs.

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Figure 1: Nes -GFP + cells are perivascular stromal cells targeted by the SNS, express high levels of Cxcl12 and are physically associated with HSCs.
Figure 2: Nes -GFP + cells are mesenchymal stem cells.
Figure 3: Adult nestin+ MSCs self-renew, differentiate and transfer haematopoietic activity in vivo.
Figure 4: Regulation of HSC maintenance by nestin + MSCs.

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Gene Expression Omnibus

Data deposits

The microarray data have been deposited in the Gene Expression Omnibus (GEO) databank (http://www.ncbi.nlm.nih.gov/geo) under the accession number GSE21941.

References

  1. 1

    Calvi, L. M. et al. Osteoblastic cells regulate the haematopoietic stem cell niche. Nature 425, 841–846 (2003)

  2. 2

    Zhang, J. et al. Identification of the haematopoietic stem cell niche and control of the niche size. Nature 425, 836–841 (2003)

  3. 3

    Adams, G. B. et al. Therapeutic targeting of a stem cell niche. Nature Biotechnol. 25, 238–243 (2007)

  4. 4

    Nilsson, S. K., Johnston, H. M. & Coverdale, J. A. Spatial localization of transplanted hemopoietic stem cells: inferences for the localization of stem cell niches. Blood 97, 2293–2299 (2001)

  5. 5

    Adams, G. B. et al. Stem cell engraftment at the endosteal niche is specified by the calcium-sensing receptor. Nature 439, 599–603 (2006)

  6. 6

    Arai, F. et al. Tie2/angiopoietin-1 signaling regulates hematopoietic stem cell quiescence in the bone marrow niche. Cell 118, 149–161 (2004)

  7. 7

    Nilsson, S. K. et al. Osteopontin, a key component of the hematopoietic stem cell niche and regulator of primitive hematopoietic progenitor cells. Blood 106, 1232–1239 (2005)

  8. 8

    Stier, S. et al. Osteopontin is a hematopoietic stem cell niche component that negatively regulates stem cell pool size. J. Exp. Med. 201, 1781–1791 (2005)

  9. 9

    Wilson, A. et al. c-Myc controls the balance between hematopoietic stem cell self-renewal and differentiation. Genes Dev. 18, 2747–2763 (2004)

  10. 10

    Xie, Y. et al. Detection of functional haematopoietic stem cell niche using real-time imaging. Nature 457, 97–101 (2009)

  11. 11

    Visnjic, D. et al. Hematopoiesis is severely altered in mice with an induced osteoblast deficiency. Blood 103, 3258–3264 (2004)

  12. 12

    Zhu, J. et al. Osteoblasts support B-lymphocyte commitment and differentiation from hematopoietic stem cells. Blood 109, 3706–3712 (2007)

  13. 13

    Lymperi, S. et al. Strontium can increase some osteoblasts without increasing hematopoietic stem cells. Blood 111, 1173–1181 (2008)

  14. 14

    Kiel, M. J., Yilmaz, O. H., Iwashita, T., Terhorst, C. & Morrison, S. J. SLAM family receptors distinguish hematopoietic stem and progenitor cells and reveal endothelial niches for stem cells. Cell 121, 1109–1121 (2005)

  15. 15

    Sugiyama, T., Kohara, H., Noda, M. & Nagasawa, T. Maintenance of the hematopoietic stem cell pool by CXCL12-CXCR4 chemokine signaling in bone marrow stromal cell niches. Immunity 25, 977–988 (2006)

  16. 16

    Katayama, Y. et al. Signals from the sympathetic nervous system regulate hematopoietic stem cell egress from bone marrow. Cell 124, 407–421 (2006)

  17. 17

    Méndez-Ferrer, S., Battista, M. & Frenette, P. S. Cooperation of β2- and β3-adrenergic receptors in hematopoietic progenitor cell mobilization. Ann. NY Acad. Sci. 1192, 139–144 (2010)

  18. 18

    Méndez-Ferrer, S., Lucas, D., Battista, M. & Frenette, P. S. Haematopoietic stem cell release is regulated by circadian oscillations. Nature 452, 442–447 (2008)

  19. 19

    Yamazaki, K. & Allen, T. D. Ultrastructural morphometric study of efferent nerve terminals on murine bone marrow stromal cells, and the recognition of a novel anatomical unit: the “neuro-reticular complex”. Am. J. Anat. 187, 261–276 (1990)

  20. 20

    Mignone, J. L., Kukekov, V., Chiang, A. S., Steindler, D. & Enikolopov, G. Neural stem and progenitor cells in nestin-GFP transgenic mice. J. Comp. Neurol. 469, 311–324 (2004)

  21. 21

    Sacchetti, B. et al. Self-renewing osteoprogenitors in bone marrow sinusoids can organize a hematopoietic microenvironment. Cell 131, 324–336 (2007)

  22. 22

    Crisan, M. et al. A perivascular origin for mesenchymal stem cells in multiple human organs. Cell Stem Cell 3, 301–313 (2008)

  23. 23

    Chan, C. K. et al. Endochondral ossification is required for haematopoietic stem-cell niche formation. Nature 457, 490–494 (2009)

  24. 24

    Mignone, J. L. et al. Neural potential of a stem cell population in the hair follicle. Cell Cycle 6, 2161–2170 (2007)

  25. 25

    Stemple, D. L. & Anderson, D. J. Isolation of a stem cell for neurons and glia from the mammalian neural crest. Cell 71, 973–985 (1992)

  26. 26

    Pardal, R., Ortega-Saenz, P., Duran, R. & Lopez-Barneo, J. Glia-like stem cells sustain physiologic neurogenesis in the adult mammalian carotid body. Cell 131, 364–377 (2007)

  27. 27

    Crisan, M. et al. Purification and culture of human blood vessel-associated progenitor cells. Curr. Protoc. Stem Cell Biol. 2, 2B.2.1–2B.2.13 (2008)

  28. 28

    Molofsky, A. V. et al. Bmi-1 dependence distinguishes neural stem cell self-renewal from progenitor proliferation. Nature 425, 962–967 (2003)

  29. 29

    Dacquin, R., Starbuck, M., Schinke, T. & Karsenty, G. Mouse α1(I)-collagen promoter is the best known promoter to drive efficient Cre recombinase expression in osteoblast. Dev. Dyn. 224, 245–251 (2002)

  30. 30

    Soriano, P. Generalized lacZ expression with the ROSA26 Cre reporter strain. Nature Genet. 21, 70–71 (1999)

  31. 31

    Tronche, F. et al. Disruption of the glucocorticoid receptor gene in the nervous system results in reduced anxiety. Nature Genet. 23, 99–103 (1999)

  32. 32

    Balordi, F. & Fishell, G. Mosaic removal of hedgehog signaling in the adult SVZ reveals that the residual wild-type stem cells have a limited capacity for self-renewal. J. Neurosci. 27, 14248–14259 (2007)

  33. 33

    Sousa, V. H., Miyoshi, G., Hjerling-Leffler, J., Karayannis, T. & Fishell, G. Characterization of Nkx6–2-derived neocortical interneuron lineages. Cereb. Cortex 19 (Suppl. 1). i1–i10 (2009)

  34. 34

    Nakashima, K. et al. The novel zinc finger-containing transcription factor osterix is required for osteoblast differentiation and bone formation. Cell 108, 17–29 (2002)

  35. 35

    Nagoshi, N. et al. Ontogeny and multipotency of neural crest-derived stem cells in mouse bone marrow, dorsal root ganglia, and whisker pad. Cell Stem Cell 2, 392–403 (2008)

  36. 36

    Takashima, Y. et al. Neuroepithelial cells supply an initial transient wave of MSC differentiation. Cell 129, 1377–1388 (2007)

  37. 37

    Semerad, C. L. et al. G-CSF potently inhibits osteoblast activity and CXCL12 mRNA expression in the bone marrow. Blood 106, 3020–3027 (2005)

  38. 38

    Mendez-Ferrer, S. & Frenette, P. S. Hematopoietic stem cell trafficking: regulated adhesion and attraction to bone marrow microenvironment. Ann. NY Acad. Sci. 1116, 392–413 (2007)

  39. 39

    Buch, T. et al. A Cre-inducible diphtheria toxin receptor mediates cell lineage ablation after toxin administration. Nature Methods 2, 419–426 (2005)

  40. 40

    Lo Celso, C. et al. Live-animal tracking of individual haematopoietic stem/progenitor cells in their niche. Nature 457, 92–96 (2009)

  41. 41

    Friedenstein, A. J., Chailakhjan, R. K. & Lalykina, K. S. The development of fibroblast colonies in monolayer cultures of guinea-pig bone marrow and spleen cells. Cell Tissue Kinet. 3, 393–403 (1970)

  42. 42

    Katayama, Y. et al. PSGL-1 participates in E-selectin-mediated progenitor homing to bone marrow: evidence for cooperation between E-selectin ligands and alpha4 integrin. Blood 102, 2060–2067 (2003)

  43. 43

    Berger, S. I., Posner, J. M. & Ma’ayan, A. Genes2Networks: connecting lists of gene symbols using mammalian protein interactions databases. BMC Bioinform. 8, 372 (2007)

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Acknowledgements

We thank M. García-Fernández, Y. Kunisaki, C. Scheiermann, J. Isern, E. Nistal-Villan and D. Lucas for help with some experiments; M. Kiel and S. Morrison for advice about immunohistological analysis of HSCs; C. Lin for help with intravital microscopy imaging; G. Fishell for the gift of Nes-creERT2 and RCE:loxP transgenic mice; J. Ahmed, W. Kao and J. Godbold for help with immunofluorescence and statistical analyses; S. Lymperi for advice about LT-CIC; L. Silberstein, G. Khitrov and W. Zhang for help with microarray experiments; M. Grisotto for help with cell sorting; and L. Shang, A. J. Peired and C. Prophete for help with animals. This work was supported by the National Institutes of Health (R01 grants DK056638, HL69438, HL097819) and the Department of Defence (Idea Development Award PC060271) to P.S.F. and by the National Institute of Mental Health and Ira Hazan Fund to G.N.E. S.M.-F. is the recipient of a Scholar Award by the American Society of Hematology. P.S.F. is an Established Investigator of the American Heart Association.

Author information

All authors contributed to the design of experiments. S.M.-F. performed experiments, analysed data and wrote the manuscript. T.V.M. and S.M.-F. performed experiments involving depletion of nestin+ cells and lineage tracing studies. F.F. performed intravital homing experiments. A.M.’a., A.R.M. and B.D.M. designed and performed analyses of microarray experiments. G.N.E., D.T.S. and S.A.L. contributed reagents and provided advice on the manuscript. P.S.F. supervised experiments and the overall study, and wrote the manuscript.

Correspondence to Simón Méndez-Ferrer or Paul S. Frenette.

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Méndez-Ferrer, S., Michurina, T., Ferraro, F. et al. Mesenchymal and haematopoietic stem cells form a unique bone marrow niche. Nature 466, 829–834 (2010) doi:10.1038/nature09262

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