Liver sinusoidal endothelial cells (LSEC) isolation from rat's liver

Saharudin, Shahida and A. Talib, Norlelawati and Abdullah, Nor Zamzila and Ab Rahman, Jamalludin and Buyong, Zunariah (2017) Liver sinusoidal endothelial cells (LSEC) isolation from rat's liver. In: 2nd International Anatomical and Biomedical Scientific Conference 2017 (IABS), 1st-2nd August 2017, Serdang, Selangor. (Unpublished)

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Abstract

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 handling cells ensure high cell yield, minimum morphology disruption and cell contamination. We presented a method of liver perfusion procedure followed by the isolation of LSEC. Seven Sprague Dawley rats underwent these procedures under anaesthesia. Liver perfusion was done as previously described. Briefly, LSECs 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 were evaluated by light and electron microscopy. 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 g rat. After 8 hours of culture, LSEC monolayers were contaminated with less than 5% of other cells. This method is reliable and reproducible for isolation of LSEC to enable the study of the structure and function of these cells in vitro. However, an improvement in the perfusion skills and isolation technique is vital to ensure better cell purity.

Item Type:	Conference or Workshop Item (Poster)
Additional Information:	5178/83933
Uncontrolled Keywords:	Liver Sinusoidal Endothelial Cells, Isolation, Perfusion
Subjects:	R Medicine > R Medicine (General)
Kulliyyahs/Centres/Divisions/Institutes (Can select more than one option. Press CONTROL button):	Kulliyyah of Medicine Kulliyyah of Medicine > Department of Basic Medical Kulliyyah of Medicine > Department of Community Medicine (Effective: 1st January 2011) Kulliyyah of Medicine > Department of Pathology & Lab Medicine
Depositing User:	DR ZUNARIAH BUYONG
Date Deposited:	28 Oct 2020 14:58
Last Modified:	28 Oct 2020 14:58
URI:	http://irep.iium.edu.my/id/eprint/83933

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