open access

Vol 27, No 4 (2021)
Research paper
Published online: 2022-01-28
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Characteristic of cells isolated from human Abdominal Aortic Aneurysm samples cultured in vitro

DOI: 10.5603/AA.2021.0014
·
Acta Angiologica 2021;27(4):120-129.
Affiliations
  1. Medical University of Silesia, Department Of Molecular Biology, Medyków 18, C1/IV p., 40-752 Katowice, Poland
  2. Medical University of Silesia, Department of Medical Genetics, Medyków 18, C1/IV p., 40-752 Katowice, Poland
  3. Medical University of Silesia, Department of General Vascular Surgery, Ziołowa 45/47, 40-635 Katowice, Poland
  4. Department of Surgical Oncology and Vascular Surgery, Oncological Centre in Katowice, ul. Raciborska 26, 40-074 Katowice, Poland

open access

Vol 27, No 4 (2021)
-- For assignment --
Published online: 2022-01-28

Abstract

Abstract Background: This study aimed to standardize cell culture methods for major cell types isolated from three layers of human AAA. We also aimed to determine cell types in each layer of each AAA segment and compare them with cell types in layers of control, unchanged segments. Material and methods We divided AAAs into three segments along the AAA and control segments flanking the aneurysm. Isolated cells following expansion were analyzed by flow cytometry, immunochemistry and microscopic methods. Fluorochrome-conjugated antibodies were used to detect the three major cell types (endothelial cells, smooth muscle cells, and fibroblasts) in each layer of every AAA segment. Results: Culture of cells from the three AAA segments was successfully established in 21% of patients. In all of the layers, only a small proportion of cells showed layer- specific markers of cell types. The majority of cells from every layer were positive for CD90, which is considered specific marker of fibroblasts in the aorta. Conclusions: We describe methodology for isolation of cells, their culture conditions, and phenotypic characterization for AAA. The wall of AAA loses its specific types of cells in all of the layers compared with the normal abdominal aortic wall.

Abstract

Abstract Background: This study aimed to standardize cell culture methods for major cell types isolated from three layers of human AAA. We also aimed to determine cell types in each layer of each AAA segment and compare them with cell types in layers of control, unchanged segments. Material and methods We divided AAAs into three segments along the AAA and control segments flanking the aneurysm. Isolated cells following expansion were analyzed by flow cytometry, immunochemistry and microscopic methods. Fluorochrome-conjugated antibodies were used to detect the three major cell types (endothelial cells, smooth muscle cells, and fibroblasts) in each layer of every AAA segment. Results: Culture of cells from the three AAA segments was successfully established in 21% of patients. In all of the layers, only a small proportion of cells showed layer- specific markers of cell types. The majority of cells from every layer were positive for CD90, which is considered specific marker of fibroblasts in the aorta. Conclusions: We describe methodology for isolation of cells, their culture conditions, and phenotypic characterization for AAA. The wall of AAA loses its specific types of cells in all of the layers compared with the normal abdominal aortic wall.

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Keywords

Abdominal aortic aneurysm; aortic adventitial fibroblast; aortic endothelial cells; aorta smooth muscles; AAA segmentation

About this article
Title

Characteristic of cells isolated from human Abdominal Aortic Aneurysm samples cultured in vitro

Journal

Acta Angiologica

Issue

Vol 27, No 4 (2021)

Article type

Research paper

Pages

120-129

Published online

2022-01-28

Page views

1070

Article views/downloads

123

DOI

10.5603/AA.2021.0014

Bibliographic record

Acta Angiologica 2021;27(4):120-129.

Keywords

Abdominal aortic aneurysm
aortic adventitial fibroblast
aortic endothelial cells
aorta smooth muscles
AAA segmentation

References (33)
  1. Davis FM, Rateri DL, Daugherty A. Abdominal aortic aneurysm: novel mechanisms and therapies. Curr Opin Cardiol. 2015; 30(6): 566–573.
  2. Sidloff D, Stather P, Dattani N, et al. Aneurysm global epidemiology study: public health measures can further reduce abdominal aortic aneurysm mortality. Circulation. 2014; 129(7): 747–753.
  3. Humphrey JD, Holzapfel GA. Mechanics, mechanobiology, and modeling of human abdominal aorta and aneurysms. J Biomech. 2012; 45(5): 805–814.
  4. Vliet JA, Boll A. Abdominal aortic aneurysm. The Lancet. 1997; 349(9055): 863–866.
  5. Kuivaniemi H, Ryer EJ, Elmore JR, et al. Understanding the pathogenesis of abdominal aortic aneurysms. Expert Rev Cardiovasc Ther. 2015; 13(9): 975–987.
  6. Mai J, Virtue A, Shen J, et al. An evolving new paradigm: endothelial cells--conditional innate immune cells. J Hematol Oncol. 2013; 6: 61.
  7. Riches K, Angelini TG, Mudhar GS, et al. Exploring smooth muscle phenotype and function in a bioreactor model of abdominal aortic aneurysm. J Transl Med. 2013; 11: 208.
  8. Jana S, Hu M, Shen M, et al. Extracellular matrix, regional heterogeneity of the aorta, and aortic aneurysm. Exp Mol Med. 2019; 51(12): 1–15.
  9. Karnik SK, Brooke BS, Bayes-Genis A, et al. A critical role for elastin signaling in vascular morphogenesis and disease. Development. 2003; 130(2): 411–423.
  10. Lesiak M, Augusciak-Duma A, Stepien KL, et al. Searching for new molecular markers for cells obtained from abdominal aortic aneurysm. J Appl Genet. 2021; 62(3): 487–497.
  11. Moll FL, Powell JT, Fraedrich G, et al. European Society for Vascular Surgery. Management of abdominal aortic aneurysms clinical practice guidelines of the European society for vascular surgery. Eur J Vasc Endovasc Surg. 2011; 41 Suppl 1: S1–S58.
  12. Visonà SD, de Boer OJ, Mackaaij C, et al. Immunophenotypic analysis of the chronological events of tissue repair in aortic medial dissections. Cardiovasc Pathol. 2018; 34: 9–14.
  13. Tanaskovic I, Ilic S, Jurisic V, et al. Histochemical, immunohistochemical and ultrastructural analysis of aortic wall in neonatal coarctation. Rom J Morphol Embryol. 2019; 60(4): 1291–1298.
  14. Yuan SM, Wu N. Aortic α-smooth muscle actin expressions in aortic disorders and coronary artery disease: An immunohistochemical study. Anatol J Cardiol. 2018; 19(1): 11–16.
  15. Müller AM, Hermanns MI, Skrzynski C, et al. Expression of the endothelial markers PECAM-1, vWf, and CD34 in vivo and in vitro. Exp Mol Pathol. 2002; 72(3): 221–229.
  16. Lertkiatmongkol P, Liao D, Mei H, et al. Endothelial functions of platelet/endothelial cell adhesion molecule-1 (CD31). Curr Opin Hematol. 2016; 23(3): 253–259.
  17. Nakada MT, Amin K, Christofidou-Solomidou M, et al. Antibodies against the first Ig-like domain of human platelet endothelial cell adhesion molecule-1 (PECAM-1) that inhibit PECAM-1-dependent homophilic adhesion block in vivo neutrophil recruitment. J Immunol. 2000; 164(1): 452–462.
  18. De Jong A, Eikenboom J. Developments in the diagnostic procedures for von Willebrand disease. J Thromb Haemost. 2016; 14(3): 449–460.
  19. Valentijn KM, Eikenboom J. Weibel-Palade bodies: a window to von Willebrand disease. J Thromb Haemost. 2013; 11(4): 581–592.
  20. Komarova YA, Kruse K, Mehta D, et al. Protein Interactions at Endothelial Junctions and Signaling Mechanisms Regulating Endothelial Permeability. Circ Res. 2017; 120(1): 179–206.
  21. Shi ZD, Tarbell JM. Fluid flow mechanotransduction in vascular smooth muscle cells and fibroblasts. Ann Biomed Eng. 2011; 39(6): 1608–1619.
  22. Yuan SM. α-Smooth Muscle Actin and ACTA2 Gene Expressions in Vasculopathies. Braz J Cardiovasc Surg. 2015; 30(6): 644–649.
  23. Owens GK, Kumar MS, Wamhoff BR. Molecular regulation of vascular smooth muscle cell differentiation in development and disease. Physiol Rev. 2004; 84(3): 767–801.
  24. Pyo R, Lee JK, Shipley JM, et al. Targeted gene disruption of matrix metalloproteinase-9 (gelatinase B) suppresses development of experimental abdominal aortic aneurysms. J Clin Invest. 2000; 105(11): 1641–1649.
  25. Hasan DM, Mahaney KB, Magnotta VA, et al. Macrophage imaging within human cerebral aneurysms wall using ferumoxytol-enhanced MRI: a pilot study. Arterioscler Thromb Vasc Biol. 2012; 32(4): 1032–1038.
  26. Aoki T, Kataoka H, Morimoto M, et al. Macrophage-derived matrix metalloproteinase-2 and -9 promote the progression of cerebral aneurysms in rats. Stroke. 2007; 38(1): 162–169.
  27. Augusciak-Duma A, Stepien KL, Lesiak M, et al. Expression gradient of metalloproteinases and their inhibitors from proximal to distal segments of abdominal aortic aneurysm. J Appl Genet. 2021; 62(3): 499–506.
  28. Rege TA, Hagood JS. Thy-1, a versatile modulator of signaling affecting cellular adhesion, proliferation, survival, and cytokine/growth factor responses. Biochim Biophys Acta. 2006; 1763(10): 991–999.
  29. Craig W, Kay R, Cutler RL, et al. Expression of Thy-1 on human hematopoietic progenitor cells. J Exp Med. 1993; 177(5): 1331–1342.
  30. Prockop DJ, Kivirikko KI. Collagens: molecular biology, diseases, and potentials for therapy. Annu Rev Biochem. 1995; 64: 403–434.
  31. Stenmark KR, Yeager ME, El Kasmi KC, et al. The adventitia: essential regulator of vascular wall structure and function. Annu Rev Physiol. 2013; 75: 23–47.
  32. Lammers S, Scott D, Hunter K, et al. Mechanics and Function of the Pulmonary Vasculature: Implications for Pulmonary Vascular Disease and Right Ventricular Function. Compr Physiol. 2012; 2(1): 295–319.
  33. Ziaja, D. Drożność tętnicy krezkowej dolnej, odwarstwienie skrzepliny oraz rozległość tętniaka jako kliniczny marker nasilenia zapalenia i ekspresji cytokin, protein adaptorowych oraz zawartość metali i niemetali w ścianie podnerkowego tętniaka aorty brzusznej. PhD Thesis, Medical University of Silesia, Poland. 2013.

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