Endothelial and perivascular cells maintain haematopoietic stem cells - PubMed (original) (raw)
Endothelial and perivascular cells maintain haematopoietic stem cells
Lei Ding et al. Nature. 2012.
Abstract
Several cell types have been proposed to create niches for haematopoietic stem cells (HSCs). However, the expression patterns of HSC maintenance factors have not been systematically studied and no such factor has been conditionally deleted from any candidate niche cell. Thus, the cellular sources of these factors are undetermined. Stem cell factor (SCF; also known as KITL) is a key niche component that maintains HSCs. Here, using Scf(gfp) knock-in mice, we found that Scf was primarily expressed by perivascular cells throughout the bone marrow. HSC frequency and function were not affected when Scf was conditionally deleted from haematopoietic cells, osteoblasts, nestin-cre- or nestin-creER-expressing cells. However, HSCs were depleted from bone marrow when Scf was deleted from endothelial cells or leptin receptor (Lepr)-expressing perivascular stromal cells. Most HSCs were lost when Scf was deleted from both endothelial and Lepr-expressing perivascular cells. Thus, HSCs reside in a perivascular niche in which multiple cell types express factors that promote HSC maintenance.
Conflict of interest statement
The authors declare no competing financial interests.
Figures
Figure 1. Scfgfp is a strong loss-of-function allele and Scf is primarily expressed by perivascular cells in the bone marrow
a and b, Scfgfp/gfp homozygous mice died perinatally and were severely anemic (n=4-20). c, Scf transcripts in livers from newborn mice by qRT-PCR (n=3). d and e, Newborn liver cellularity and HSC frequency (n=4). f, Irradiated mice (CD45.1+) were transplanted with 3×105 newborn liver cells from Scfgfp/gfp, Scfgfp/+ or Scf+/+ donor (CD45.2+) mice along with 3×105 recipient (CD45.1+) bone marrow cells (3-4 experiments with 13-18 mice/genotype). g, Scf-GFP was expressed by rare non-haematopoietic stromal cells (n=8). h-j, GFP was primarily expressed by perivascular cells in the bone marrow of Scfgfp/+ mice. Endothelial cells were stained with an anti-Endoglin antibody. k-n, GFP was not detected in bone-lining osteoblast lineage cells (Osteopontin) in the diaphysis (k-m) or in trabecular bone (n). o-q, Higher magnification images of a sinusoid. r-u, A CD150+CD48-Lineage- candidate HSC (arrow) localized adjacent to a GFP-expressing perivascular cell. Nuclei were stained with DAPI (in blue). All data represent mean±s.d. Two-tail student’s t-tests were used to assess statistical significance: ** p<0.01, *** p<0.001. Scale bars in (j), (m) and (n) are 50um. Scale bars in (q) and (u) are 20um.
Figure 2. Scf is required for adult HSC maintenance
a, Homozygous Scf-/- mutant mice generated from germline recombination of the Scffl allele were perinatal lethal and anemic. b, Scf transcripts amplified by RT-PCR from the livers of newborn mice. c, Global deletion of Scf in Ubc-CreER; Scffl/fl mice led to anemia (n=5-6). d and e, Global deletion of Scf in adult mice significantly reduced cellularity and HSC frequency in bone marrow (two femurs and two tibias) and spleen (n=8-10). f, To perform a limit dilution analysis, three doses of donor bone marrow cells were competitively transplanted into irradiated mice. ELDA software (
http://bioinf.wehi.edu.au/software/elda/
) was used to calculate HSC frequency and statistical significance (two experiments). g, 3×105 donor bone marrow cells were transplanted with 3×105 recipient bone marrow cells into irradiated recipient mice (3 experiments with a total of 12-14 recipients/genotype). h, HSCs did not express _Scf-G_FP by flow cytometry. Δ, recombined Scffl allele; +, wild-type allele of Scf.
Figure 3. SCF from haematopoietic cells, osteoblasts, and _Nestin-Cre-_expressing stromal cells is not required for HSC maintenance
a, Vav1-Cre recombined the loxpEYFP reporter in virtually all HSCs, CD45+, and Ter119+ haematopoietic cells. b and c, Deletion of Scf from haematopoietic cells did not significantly affect bone marrow or spleen cellularity or HSC frequency (n=4). d, A competitive reconstitution assay with Vav-1-Cre; Scffl/-, Scf+/- and Scf+/+ bone marrow cells (two experiments with a total of 10 recipients/genotype). e, Col2.3-Cre recombined the loxpEYFP reporter in bone-lining osteoblast lineage cells. f, Nestin-Cre recombined the loxpEYFP reporter in rare stromal cells around larger blood vessels. g, Bone marrow and spleen cellularity and h, HSC frequency in Col2.3-Cre; Scffl/- mice relative to controls (n=5-6). i, A competitive reconstitution assay with Col2.3-Cre; Scffl/-, Scf+/- and Scf+/+ bone marrow cells (3-5 experiments with a total of 14-22 recipients/genotype). j, Bone marrow and spleen cellularity and k, HSC frequency in Nestin-Cre; Scffl/- mice relative to controls (n=5-7). l, 3×105 donor bone marrow cells from Nestin-Cre; Scffl/- and Scf+/+ mice gave similar levels of donor cell reconstitution in irradiated mice. Reconstitution levels from Scf+/- cells were modestly but significantly lower (3-5 experiments with a total of 14-24 recipient mice/genotype). Δ, recombined Scffl allele; +, wild-type allele; −, germline deleted allele. NS, not significant. Scale bar is 100um in (e) and 50um in (f).
Figure 4. Deletion of Scf from endothelial cells depletes HSCs
a and b, Tie2-Cre recombined the loxpEYFP conditional reporter in VE-cadherin+ endothelial cells and in haematopoietic cells in the bone marrow. c, Bone marrow and spleen cellularity in Tie2-Cre; Scffl/- mice and littermate controls (n=4-7). d, HSC frequency in Tie2-Cre; Scffl/- mice and controls (n=4-7). e, HSC frequency in the liver of newborn Tie2-Cre; Scffl/- mice and controls (n=3-6). f, HSC frequency in one month-old Tie2-Cre; Scffl/- mice and controls (n=3-4). g, Bone marrow cells from Tie2-Cre; Scffl/- mice gave significantly lower levels of reconstitution relative to cells from Scf+/- and Scf+/+ mice (3-5 experiments with a total of 15-25 recipients/genotype).
Figure 5. Deletion of Scf from _Lepr-Cre_-expressing perivascular stromal cells depletes HSCs in the bone marrow
a-c, Lepr-Cre recombined the loxpEYFP reporter in perisinusoidal stromal cells in the bone marrow but not in bone-lining or haematopoietic cells. d, Lepr-Cre did not recombine in VE-cadherin+ endothelial cells. e, 0.013±0.009% (mean±s.d.; n=3) of bone marrow cells from Lepr-Cre; loxpEYFP mice were EYFP+. f, Spleen size and g, bone marrow and spleen cellularity (n=4-7). h, HSC frequency (n=4-7). i, Total HSC numbers (including bone marrow and spleen) in Lepr-Cre; Scffl/gfp mice (n=4-7). j, Limit dilution analysis of the frequency of long-term multilineage reconstituting cells in the bone marrow of Lepr-Cre; Scffl/gfp mice relative to controls (two experiments). k, HSC frequency in the newborn liver (n=4-11). l, HSC frequency in one month-old Lepr-Cre; Scffl/gfp mice and controls (n=3-6). m, Tie2-Cre; Lepr-Cre; Scffl/- mice had significantly reduced bone marrow cellularity and increased spleen cellularity compared to Scf+/- or Tie2-Cre; Scffl/- controls (n=4-11). n, Deletion of Scf from endothelial and perivascular stromal cells in Tie2-Cre; Lepr-Cre; Scffl/- mice greatly depleted HSCs from adult bone marrow (n=4-11). o, Total HSC number was significantly reduced in Tie2-Cre; Lepr-Cre; Scffl/- mice compared to Tie2-Cre; Scffl/- or Lepr-Cre; Scffl/- mice (n=4-11). Scale bars are 50um.
Comment in
- Stem cells: The right neighbour.
Shestopalov IA, Zon LI. Shestopalov IA, et al. Nature. 2012 Jan 25;481(7382):453-5. doi: 10.1038/481453a. Nature. 2012. PMID: 22281591 No abstract available. - This niche is a maze; an amazing niche.
Hanoun M, Frenette PS. Hanoun M, et al. Cell Stem Cell. 2013 Apr 4;12(4):391-2. doi: 10.1016/j.stem.2013.03.012. Cell Stem Cell. 2013. PMID: 23561440 Free PMC article.
Similar articles
- Haematopoietic stem cells and early lymphoid progenitors occupy distinct bone marrow niches.
Ding L, Morrison SJ. Ding L, et al. Nature. 2013 Mar 14;495(7440):231-5. doi: 10.1038/nature11885. Epub 2013 Feb 24. Nature. 2013. PMID: 23434755 Free PMC article. - Mesenchymal and haematopoietic stem cells form a unique bone marrow niche.
Méndez-Ferrer S, Michurina TV, Ferraro F, Mazloom AR, Macarthur BD, Lira SA, Scadden DT, Ma'ayan A, Enikolopov GN, Frenette PS. Méndez-Ferrer S, et al. Nature. 2010 Aug 12;466(7308):829-34. doi: 10.1038/nature09262. Nature. 2010. PMID: 20703299 Free PMC article. - CXCL12 in early mesenchymal progenitors is required for haematopoietic stem-cell maintenance.
Greenbaum A, Hsu YM, Day RB, Schuettpelz LG, Christopher MJ, Borgerding JN, Nagasawa T, Link DC. Greenbaum A, et al. Nature. 2013 Mar 14;495(7440):227-30. doi: 10.1038/nature11926. Epub 2013 Feb 24. Nature. 2013. PMID: 23434756 Free PMC article. - Pericytes, integral components of adult hematopoietic stem cell niches.
Sá da Bandeira D, Casamitjana J, Crisan M. Sá da Bandeira D, et al. Pharmacol Ther. 2017 Mar;171:104-113. doi: 10.1016/j.pharmthera.2016.11.006. Epub 2016 Nov 28. Pharmacol Ther. 2017. PMID: 27908803 Review. - [Bone and Stem Cells. Bone marrow microenvironment niches for hematopoietic stem and progenitor cells].
Nagasawa T. Nagasawa T. Clin Calcium. 2014 Apr;24(4):517-26. Clin Calcium. 2014. PMID: 24681497 Review. Japanese.
Cited by
- Skeletal stem and progenitor cells in bone physiology, ageing and disease.
Melis S, Trompet D, Chagin AS, Maes C. Melis S, et al. Nat Rev Endocrinol. 2024 Oct 8. doi: 10.1038/s41574-024-01039-y. Online ahead of print. Nat Rev Endocrinol. 2024. PMID: 39379711 Review. - The role of immune cells settled in the bone marrow on adult hematopoietic stem cells.
Xu H, Li Y, Gao Y. Xu H, et al. Cell Mol Life Sci. 2024 Oct 5;81(1):420. doi: 10.1007/s00018-024-05445-3. Cell Mol Life Sci. 2024. PMID: 39367881 Free PMC article. Review. - Cytomegalovirus results in poor graft function via bone marrow-derived endothelial progenitor cells.
Lv W, Zhou Y, Zhao K, Xuan L, Huang F, Fan Z, Chang Y, Yi Z, Jin H, Liang Y, Liu Q. Lv W, et al. Front Microbiol. 2024 Sep 18;15:1463335. doi: 10.3389/fmicb.2024.1463335. eCollection 2024. Front Microbiol. 2024. PMID: 39360328 Free PMC article. - Autophagy as a Guardian of Vascular Niche Homeostasis.
Dergilev K, Gureenkov A, Parfyonova Y. Dergilev K, et al. Int J Mol Sci. 2024 Sep 20;25(18):10097. doi: 10.3390/ijms251810097. Int J Mol Sci. 2024. PMID: 39337582 Free PMC article. Review. - Engraftment of human mesenchymal stem cells in a severely immunodeficient mouse.
Kato Y, Ohno Y, Ito R, Taketani T, Matsuzaki Y, Miyagi S. Kato Y, et al. Inflamm Regen. 2024 Sep 26;44(1):40. doi: 10.1186/s41232-024-00353-2. Inflamm Regen. 2024. PMID: 39327616 Free PMC article.
References
- Kiel MJ, Morrison SJ. Uncertainty in the niches that maintain haematopoietic stem cells. Nat Rev Immunol. 2008;8:290–301. - PubMed
- Wilson A, Trumpp A. Bone-marrow haematopoietic-stem-cell niches. Nat Rev Immunol. 2006;6:93–106. - PubMed
- Arai F, et al. Tie2/angiopoietin-1 signaling regulates hematopoietic stem cell quiescence in the bone marrow niche. Cell. 2004;118:149–161. - PubMed
- Calvi LM, et al. Osteoblastic cells regulate the haematopoietic stem cell niche. Nature. 2003;425:841–846. - PubMed
Publication types
MeSH terms
Substances
Grants and funding
- R01AG040209/AG/NIA NIH HHS/United States
- R01 AG040209/AG/NIA NIH HHS/United States
- R01 HL097760-04/HL/NHLBI NIH HHS/United States
- R01 HL097760/HL/NHLBI NIH HHS/United States
- HHMI/Howard Hughes Medical Institute/United States
- 5R01HL097760/HL/NHLBI NIH HHS/United States
LinkOut - more resources
Full Text Sources
Other Literature Sources
Medical
Molecular Biology Databases
Miscellaneous