Liver Sinusoidal Endothelial Cell (LSEC) isolation following a liver perfusion technique

Introduction: Liver perfusion has been the standard method to digest and isolate liver cells including liver sinusoidal endothelial cells (LSEC). Poor cannulating skills through portal vein results in a waste of animal resource. Familiarization of both liver perfusion technique and adhering strictly...

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Main Authors:	Saharudin, Shahida, A. Talib, Norlelawati, Abdullah, Nor Zamzila, Ab Rahman, Jamalludin, Buyong, Zunariah
Format:	Article
Language:	English
Published:	Kulliyyah of Medicine, International Islamic University Malaysia 2017
Subjects:	R Medicine (General)
Online Access:	http://irep.iium.edu.my/62377/ http://irep.iium.edu.my/62377/ http://irep.iium.edu.my/62377/1/62377_Liver%20Sinusoidal%20Endothelial%20Cell%20%28LSEC%29.pdf

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spelling	iium-623772018-11-26T05:15:06Z http://irep.iium.edu.my/62377/ Liver Sinusoidal Endothelial Cell (LSEC) isolation following a liver perfusion technique Saharudin, Shahida A. Talib, Norlelawati Abdullah, Nor Zamzila Ab Rahman, Jamalludin Buyong, Zunariah R Medicine (General) Introduction: Liver perfusion has been the standard method to digest and isolate liver cells including liver sinusoidal endothelial cells (LSEC). Poor cannulating skills through portal vein results in a waste of animal resource. Familiarization of both liver perfusion technique and adhering strictly to aseptic technique during cell handling ensure high cell yield, minimum morphology disruption and cell contamination. We aimed to present a method of liver perfusion procedure followed by the isolation of LSEC. Materials and method: The study was conducted with the approval of IACUC committee. Seven Sprague Dawley rats underwent these procedures under anaesthesia. Liver perfusion was done as previously described. Briefly, LSEC were isolated by liberase enzyme perfusion of the liver, isopycnic sedimentation in a two- step Percoll gradient and selective adherence. The purification and cultivation of LSEC was evaluated by light and electron microscopy. Results: Purity and viability of LSEC after selective adherence was 80.5 ± 3.5% and ≥ 95 %, respectively. The average concentration of the cells ranged from 32 - 75 x 106 per 400 gm rat. After 8 hours of culture, LSEC monolayers were contaminated with less than 5% of other cells. Conclusion: This method is reliable and reproducible for the isolation of LSEC to enable the study of structure and function of these cells in vitro. However, improvement on the perfusion skills and isolation technique are vital to ensure better cell purity. Kulliyyah of Medicine, International Islamic University Malaysia 2017-09-15 Article PeerReviewed application/pdf en http://irep.iium.edu.my/62377/1/62377_Liver%20Sinusoidal%20Endothelial%20Cell%20%28LSEC%29.pdf Saharudin, Shahida and A. Talib, Norlelawati and Abdullah, Nor Zamzila and Ab Rahman, Jamalludin and Buyong, Zunariah (2017) Liver Sinusoidal Endothelial Cell (LSEC) isolation following a liver perfusion technique. The International Medical Journal Malaysia (IMJM), 16 (Supp. 1). p. 27. ISSN 1823-4631 http://iiumedic.net/imjm/v1/volume-16-supplementary-issue-no-1/
repository_type	Digital Repository
institution_category	Local University
institution	International Islamic University Malaysia
building	IIUM Repository
collection	Online Access
language	English
topic	R Medicine (General)
spellingShingle	R Medicine (General) Saharudin, Shahida A. Talib, Norlelawati Abdullah, Nor Zamzila Ab Rahman, Jamalludin Buyong, Zunariah Liver Sinusoidal Endothelial Cell (LSEC) isolation following a liver perfusion technique
description	Introduction: Liver perfusion has been the standard method to digest and isolate liver cells including liver sinusoidal endothelial cells (LSEC). Poor cannulating skills through portal vein results in a waste of animal resource. Familiarization of both liver perfusion technique and adhering strictly to aseptic technique during cell handling ensure high cell yield, minimum morphology disruption and cell contamination. We aimed to present a method of liver perfusion procedure followed by the isolation of LSEC. Materials and method: The study was conducted with the approval of IACUC committee. Seven Sprague Dawley rats underwent these procedures under anaesthesia. Liver perfusion was done as previously described. Briefly, LSEC were isolated by liberase enzyme perfusion of the liver, isopycnic sedimentation in a two- step Percoll gradient and selective adherence. The purification and cultivation of LSEC was evaluated by light and electron microscopy. Results: Purity and viability of LSEC after selective adherence was 80.5 ± 3.5% and ≥ 95 %, respectively. The average concentration of the cells ranged from 32 - 75 x 106 per 400 gm rat. After 8 hours of culture, LSEC monolayers were contaminated with less than 5% of other cells. Conclusion: This method is reliable and reproducible for the isolation of LSEC to enable the study of structure and function of these cells in vitro. However, improvement on the perfusion skills and isolation technique are vital to ensure better cell purity.
format	Article
author	Saharudin, Shahida A. Talib, Norlelawati Abdullah, Nor Zamzila Ab Rahman, Jamalludin Buyong, Zunariah
author_facet	Saharudin, Shahida A. Talib, Norlelawati Abdullah, Nor Zamzila Ab Rahman, Jamalludin Buyong, Zunariah
author_sort	Saharudin, Shahida
title	Liver Sinusoidal Endothelial Cell (LSEC) isolation following a liver perfusion technique
title_short	Liver Sinusoidal Endothelial Cell (LSEC) isolation following a liver perfusion technique
title_full	Liver Sinusoidal Endothelial Cell (LSEC) isolation following a liver perfusion technique
title_fullStr	Liver Sinusoidal Endothelial Cell (LSEC) isolation following a liver perfusion technique
title_full_unstemmed	Liver Sinusoidal Endothelial Cell (LSEC) isolation following a liver perfusion technique
title_sort	liver sinusoidal endothelial cell (lsec) isolation following a liver perfusion technique
publisher	Kulliyyah of Medicine, International Islamic University Malaysia
publishDate	2017
url	http://irep.iium.edu.my/62377/ http://irep.iium.edu.my/62377/ http://irep.iium.edu.my/62377/1/62377_Liver%20Sinusoidal%20Endothelial%20Cell%20%28LSEC%29.pdf
first_indexed	2023-09-18T21:28:24Z
last_indexed	2023-09-18T21:28:24Z
_version_	1777412346478592000

Liver Sinusoidal Endothelial Cell (LSEC) isolation following a liver perfusion technique

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