Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2004 May;113(9):1258-65.
doi: 10.1172/JCI19628.

Abundant progenitor cells in the adventitia contribute to atherosclerosis of vein grafts in ApoE-deficient mice

Yanhua Hu  1 , Zhongyi Zhang, Evelyn Torsney, Ali R Afzal, Fergus Davison, Bernhard Metzler, Qingbo Xu
Affiliations

Abundant progenitor cells in the adventitia contribute to atherosclerosis of vein grafts in ApoE-deficient mice

Yanhua Hu et al. J Clin Invest. 2004 May.

Abstract

Recent evidence indicates that vascular progenitor cells may be the source of smooth muscle cells (SMCs) that accumulate in atherosclerotic lesions, but the origin of these progenitor cells is unknown. To explore the possibility of vascular progenitor cells existing in adults, a variety of tissues from ApoE-deficient mice were extensively examined. Immunohistochemical staining revealed that the adventitia in aortic roots harbored large numbers of cells having stem cell markers, e.g., Sca-1(+) (21%), c-kit(+) (9%), CD34(+) (15%), and Flk1(+) cells (4%), but not SSEA-1(+) embryonic stem cells. Explanted cultures of adventitial tissues using stem cell medium displayed a heterogeneous outgrowth, for example, islands of round-shaped cells surrounded by fibroblast-like cell monolayers. Isolated Sca-1(+) cells were able to differentiate into SMCs in response to PDGF-BB stimulation in vitro. When Sca-1(+) cells carrying the LacZ gene were transferred to the adventitial side of vein grafts in ApoE-deficient mice, beta-gal(+) cells were found in atherosclerotic lesions of the intima, and these cells enhanced the development of the lesions. Thus, a large population of vascular progenitor cells existing in the adventitia can differentiate into SMCs that contribute to atherosclerosis. Our findings indicate that ex vivo expansion of these progenitor cells may have implications for cellular, genetic, and tissue engineering approaches to vascular disease.

PubMed Disclaimer

Figures

Figure 1
Figure 1
Immunohistochemical staining. Aortic roots from ApoE_/_ mice were sectioned and labeled with a variety of antibodies as indicated. Sections were developed with alkaline phosphatase anti-alkaline phosphatase (APAAP) techniques and counterstained with hematoxylin (blue). Total cells and positive cells were counted under the microscope. Arrows indicate intima and media (A and C) or examples of positive cells (D, red). (A) The aortic wall. (B) A heart section. (C) The aortic root region. A_C were labeled with Sca-1 antibody and photographed at lower magnification. (D) Sections from aortic root regions at a higher magnification. (E) The graph shows data of means (± SEM) from eight animals per group.
Figure 2
Figure 2
H&E staining of cultured adventitial and medial cells. Adventitial (A_F) and medial (G and H) tissues from aortic roots were explanted onto a slide chamber bottle and cultivated in the medium for culturing stem cells for 5 days. Cells were fixed with 4% formalin and stained with H&E. Note adipocyte-like cells (arrows), a dividing cell (arrowhead), and large nuclei (arrows with dotted lines).
Figure 3
Figure 3
Immunostaining for Sca-1+ cells isolated with microbeads. Primary cultured adventitial cells were isolated with microbeads coupled with anti_Sca-1 antibody. The isolated cells were smeared, double immunostained for Sca-1 and Lin (A_C) or Sca-1 and c-kit (D_F), and counterstained for nuclei. Note that more than 90% of cells are Sca-1+Lin_c-kit+.
Figure 4
Figure 4
Immunostaining for Sca-1+ cells isolated with FACS sorting. Primary cultured adventitial cells were labeled with anti_Sca-1 antibody. Positive stained cells were sorted by FACS. (A) A single peak of isolated Sca-1+ cells (filled graph), showing no overlap with a negative control with only secondary antibody (green). (B_D) Microscopic photographs showing that the isolated cells were smeared, immunostained, and counterstained for nuclei. Note a result similar to that shown in Figure 3.
Figure 5
Figure 5
Sca-1+ cells express SMC markers in response to PDGF-BB. Sca-1+ cells were isolated with microbeads coupled with anti_Sca-1 antibodies and cultured in stem cell medium without PDGF-BB or in DMEM supplemented for 3 days with 10% FCS in the presence of PDGF-BB (10 ng/ml). (A) RNA was isolated, and RT-PCR was performed using the primers for SMC markers. (B) Adventitial Sca-1+ cells cultivated from SM-LacZ mice were cultured for 3 days with or without PDGF-BB, and developed with X-gal. (C) For immunofluorescent staining, cells were labeled with antibodies against α-actin (red), calponin (green), and smooth muscle myosin heavy chain (red) and counterstained with Hoechst 33258 (blue). SM-MHC, smooth muscle myosin heavy chain.
Figure 6
Figure 6
Creation of chimeric mice and culture of adventitial cells. Femurs of chimeric mice with ROSA26 or wild-type bone marrow were harvested and transplanted. Sections of the aortic root from chimeric mice with ROSA26 (A) or wild-type (B) bone marrow were stained with X-gal. (C and D) Cultured adventitial cells from chimeric mice with ROSA26 bone marrow (C) or SM-LacZ mice with wild-type bone marrow (D). Cells were stimulated with PDGF-BB (10 ng/ml) for 3 days and stained with β-gal. Arrows indicate β-gal+ cells.
Figure 7
Figure 7
Sca-1+/SM-LacZ cell transfer to vein grafts in mice. Sca-1+ cells were cultivated from the adventitia of SM-LacZ/ApoE_/_ mice as described for Figure 5B. Irradiated vena cava of the mouse was removed and grafted into carotid arteries of ApoE_/_ mice. Sca-1+/SM-LacZ cells (B_D) (105) were applied onto the adventitia to envelop the vein grafts. Animals were sacrificed 2 weeks (B) or 4 weeks (A, C, and D) after surgery, and the grafted tissue fragments were sectioned and stained with X-gal. A shows a negative control without Sca-1+ cells. Arrows indicate β-gal+ cells.
Figure 8
Figure 8
Sca-1+/ROSA26 cell transfer to vein grafts in mice. Sca-1+ cells cultivated from the adventitia of ROSA26/ApoE_/_ (A) and ApoE_/_ (B) mice were stained with X-gal. Irradiated vena cava of the mouse was removed and grafted into carotid arteries of ApoE_/_ mice. Sca-1+ (C, D, and E) or Sca-1_ (F) cells (105) were applied onto the adventitia to envelop the vein grafts. Animals were sacrificed 4 weeks after surgery, and the grafted tissue fragments were sectioned and stained with X-gal. For cell counting, Hoechst counterstain was used. C and E represent the adventitial area of the grafted tissue. D and F focus on the neointimal and medial layers. Note β-gal+ cells, stained blue. Arrows indicate the border of the neointima and media. (G) The graphic data are means (± SEM) of five animals per group. *Significant difference in intimal lesions between Sca-1+ and Sca-1_ group, P < 0.001. **Significant difference in the adventitia between Sca-1+ and Sca-1_ group, P < 0.001.
See this image and copyright information in PMC

Comment in

  • Lost in transdifferentiation.
    Hoofnagle MH, Wamhoff BR, Owens GK. Hoofnagle MH, et al. J Clin Invest. 2004 May;113(9):1249-51. doi: 10.1172/JCI21761. J Clin Invest. 2004. PMID: 15124012 Free PMC article.

Similar articles

See all similar articles

Cited by

See all "Cited by" articles

References

    1. Ross R, Glomset J, Harker L. Response to injury and atherogenesis. Am. J. Pathol. 1977;86:675–684. - PMC - PubMed
    1. Saiura A, Sata M, Hirata Y, Nagai R, Makuuchi M. Circulating smooth muscle progenitor cells contribute to atherosclerosis. Nat. Med. 2001;7:382–383. - PubMed
    1. Han CI, Campbell GR, Campbell JH. Circulating bone marrow cells can contribute to neointimal formation. J. Vasc. Res. 2001;38:113–119. - PubMed
    1. Shimizu K, et al. Host bone-marrow cells are a source of donor intimal smooth-muscle-like cells in murine aortic transplant arteriopathy. Nat. Med. 2001;7:738–741. - PubMed
    1. Hillebrands JL, et al. Origin of neointimal endothelium and α-actin–positive smooth muscle cells in transplant arteriosclerosis. J. Clin. Invest. 2001;107:1411–1422. - PMC - PubMed

Publication types

MeSH terms