New method for the isolation of endothelial cells from large vessels

Forough Ataollahi, Belinda Pingguan-Murphy, Ali Moradi, Wan Abu Bakar Wan Abas, Chua Kien Hui, Noor Azuan Abu Osman

Research output: Contribution to journalArticle

5 Citations (Scopus)

Abstract

Background aims: Numerous protocols for the isolation of bovine aortic endothelial cells have been described in the previous literature. However, these protocols prevent researchers from obtaining the pure population of endothelial cells. Thus, this study aimed to develop a new and economical method for the isolation of pure endothelial cells by introducing a new strategy to the enzymatic digestion method proposed by previous researchers. Methods: With the use of this method, the lumen of a bovine aorta was filled with wash medium and the outer surface of the sample was washed with alcohol for 30 seconds. Under a laminar flow hood, the inner surface of the sample was covered with filter paper. Collagenase type II was dripped onto the filter paper as a digestion enzyme. The digestion fluid was seeded in T25 flasks and fed with complete medium every 3 days. Results: The isolated cells were characterized by markers such as CD31, von Willebrand factor, 1,1'-dioctadecyl-1,3,3,3',3'-tetramethylindocarbocyanine perchlorate acetylated low-density lipoprotein and angiogenesis behavior. The purity of endothelial cells was detected by flow cytometry to be of nearly 90% purity; these results were confirmed by immunostaining. Moreover, endothelial cells formed blood vessel-like tubes in a three-dimensional environment, which is specific dynamic behavior for endothelial cells. Conclusions: The new strategy presented in the current report enables isolation of a highly pure population of endothelial cells that can survive long-term culture without inducing an overgrowth of fibroblast cells.

Original languageEnglish
Pages (from-to)1145-1152
Number of pages8
JournalCytotherapy
Volume16
Issue number8
DOIs
Publication statusPublished - 2014

Fingerprint

Endothelial Cells
Digestion
Research Personnel
von Willebrand Factor
Collagenases
Population
Blood Vessels
Aorta
Flow Cytometry
Fibroblasts
Alcohols
Enzymes

Keywords

  • Bovine aortic endothelial cells
  • Characterization
  • Fibroblast contamination
  • Filter paper
  • New strategy
  • Pure population

ASJC Scopus subject areas

  • Immunology
  • Immunology and Allergy
  • Oncology
  • Genetics(clinical)
  • Transplantation
  • Cancer Research
  • Cell Biology
  • Medicine(all)

Cite this

Ataollahi, F., Pingguan-Murphy, B., Moradi, A., Bakar Wan Abas, W. A., Kien Hui, C., & Abu Osman, N. A. (2014). New method for the isolation of endothelial cells from large vessels. Cytotherapy, 16(8), 1145-1152. https://doi.org/10.1016/j.jcyt.2014.01.010

New method for the isolation of endothelial cells from large vessels. / Ataollahi, Forough; Pingguan-Murphy, Belinda; Moradi, Ali; Bakar Wan Abas, Wan Abu; Kien Hui, Chua; Abu Osman, Noor Azuan.

In: Cytotherapy, Vol. 16, No. 8, 2014, p. 1145-1152.

Research output: Contribution to journalArticle

Ataollahi, F, Pingguan-Murphy, B, Moradi, A, Bakar Wan Abas, WA, Kien Hui, C & Abu Osman, NA 2014, 'New method for the isolation of endothelial cells from large vessels', Cytotherapy, vol. 16, no. 8, pp. 1145-1152. https://doi.org/10.1016/j.jcyt.2014.01.010
Ataollahi F, Pingguan-Murphy B, Moradi A, Bakar Wan Abas WA, Kien Hui C, Abu Osman NA. New method for the isolation of endothelial cells from large vessels. Cytotherapy. 2014;16(8):1145-1152. https://doi.org/10.1016/j.jcyt.2014.01.010
Ataollahi, Forough ; Pingguan-Murphy, Belinda ; Moradi, Ali ; Bakar Wan Abas, Wan Abu ; Kien Hui, Chua ; Abu Osman, Noor Azuan. / New method for the isolation of endothelial cells from large vessels. In: Cytotherapy. 2014 ; Vol. 16, No. 8. pp. 1145-1152.
@article{14b1329c30e445e18b2842be09640095,
title = "New method for the isolation of endothelial cells from large vessels",
abstract = "Background aims: Numerous protocols for the isolation of bovine aortic endothelial cells have been described in the previous literature. However, these protocols prevent researchers from obtaining the pure population of endothelial cells. Thus, this study aimed to develop a new and economical method for the isolation of pure endothelial cells by introducing a new strategy to the enzymatic digestion method proposed by previous researchers. Methods: With the use of this method, the lumen of a bovine aorta was filled with wash medium and the outer surface of the sample was washed with alcohol for 30 seconds. Under a laminar flow hood, the inner surface of the sample was covered with filter paper. Collagenase type II was dripped onto the filter paper as a digestion enzyme. The digestion fluid was seeded in T25 flasks and fed with complete medium every 3 days. Results: The isolated cells were characterized by markers such as CD31, von Willebrand factor, 1,1'-dioctadecyl-1,3,3,3',3'-tetramethylindocarbocyanine perchlorate acetylated low-density lipoprotein and angiogenesis behavior. The purity of endothelial cells was detected by flow cytometry to be of nearly 90{\%} purity; these results were confirmed by immunostaining. Moreover, endothelial cells formed blood vessel-like tubes in a three-dimensional environment, which is specific dynamic behavior for endothelial cells. Conclusions: The new strategy presented in the current report enables isolation of a highly pure population of endothelial cells that can survive long-term culture without inducing an overgrowth of fibroblast cells.",
keywords = "Bovine aortic endothelial cells, Characterization, Fibroblast contamination, Filter paper, New strategy, Pure population",
author = "Forough Ataollahi and Belinda Pingguan-Murphy and Ali Moradi and {Bakar Wan Abas}, {Wan Abu} and {Kien Hui}, Chua and {Abu Osman}, {Noor Azuan}",
year = "2014",
doi = "10.1016/j.jcyt.2014.01.010",
language = "English",
volume = "16",
pages = "1145--1152",
journal = "Cytotherapy",
issn = "1465-3249",
publisher = "Informa Healthcare",
number = "8",

}

TY - JOUR

T1 - New method for the isolation of endothelial cells from large vessels

AU - Ataollahi, Forough

AU - Pingguan-Murphy, Belinda

AU - Moradi, Ali

AU - Bakar Wan Abas, Wan Abu

AU - Kien Hui, Chua

AU - Abu Osman, Noor Azuan

PY - 2014

Y1 - 2014

N2 - Background aims: Numerous protocols for the isolation of bovine aortic endothelial cells have been described in the previous literature. However, these protocols prevent researchers from obtaining the pure population of endothelial cells. Thus, this study aimed to develop a new and economical method for the isolation of pure endothelial cells by introducing a new strategy to the enzymatic digestion method proposed by previous researchers. Methods: With the use of this method, the lumen of a bovine aorta was filled with wash medium and the outer surface of the sample was washed with alcohol for 30 seconds. Under a laminar flow hood, the inner surface of the sample was covered with filter paper. Collagenase type II was dripped onto the filter paper as a digestion enzyme. The digestion fluid was seeded in T25 flasks and fed with complete medium every 3 days. Results: The isolated cells were characterized by markers such as CD31, von Willebrand factor, 1,1'-dioctadecyl-1,3,3,3',3'-tetramethylindocarbocyanine perchlorate acetylated low-density lipoprotein and angiogenesis behavior. The purity of endothelial cells was detected by flow cytometry to be of nearly 90% purity; these results were confirmed by immunostaining. Moreover, endothelial cells formed blood vessel-like tubes in a three-dimensional environment, which is specific dynamic behavior for endothelial cells. Conclusions: The new strategy presented in the current report enables isolation of a highly pure population of endothelial cells that can survive long-term culture without inducing an overgrowth of fibroblast cells.

AB - Background aims: Numerous protocols for the isolation of bovine aortic endothelial cells have been described in the previous literature. However, these protocols prevent researchers from obtaining the pure population of endothelial cells. Thus, this study aimed to develop a new and economical method for the isolation of pure endothelial cells by introducing a new strategy to the enzymatic digestion method proposed by previous researchers. Methods: With the use of this method, the lumen of a bovine aorta was filled with wash medium and the outer surface of the sample was washed with alcohol for 30 seconds. Under a laminar flow hood, the inner surface of the sample was covered with filter paper. Collagenase type II was dripped onto the filter paper as a digestion enzyme. The digestion fluid was seeded in T25 flasks and fed with complete medium every 3 days. Results: The isolated cells were characterized by markers such as CD31, von Willebrand factor, 1,1'-dioctadecyl-1,3,3,3',3'-tetramethylindocarbocyanine perchlorate acetylated low-density lipoprotein and angiogenesis behavior. The purity of endothelial cells was detected by flow cytometry to be of nearly 90% purity; these results were confirmed by immunostaining. Moreover, endothelial cells formed blood vessel-like tubes in a three-dimensional environment, which is specific dynamic behavior for endothelial cells. Conclusions: The new strategy presented in the current report enables isolation of a highly pure population of endothelial cells that can survive long-term culture without inducing an overgrowth of fibroblast cells.

KW - Bovine aortic endothelial cells

KW - Characterization

KW - Fibroblast contamination

KW - Filter paper

KW - New strategy

KW - Pure population

UR - http://www.scopus.com/inward/record.url?scp=84903820840&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84903820840&partnerID=8YFLogxK

U2 - 10.1016/j.jcyt.2014.01.010

DO - 10.1016/j.jcyt.2014.01.010

M3 - Article

VL - 16

SP - 1145

EP - 1152

JO - Cytotherapy

JF - Cytotherapy

SN - 1465-3249

IS - 8

ER -