Monolayer culture expansion of Annulus fibrosus cells for intervertebral disc regeneration

Objectives/Research Problem: Tissue engineering (TE) provides an alternative approach in regenerating human tissues for clinical transplantation and drugs development purposes. In principle, TE requires good cell source, compatible biomaterial scaffold and proper signalling factors. Regeneration of...

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Main Authors:	Mohamad, Mohd Yusof, Mohamed Amin, Muhammad Azri Ifwat, Zulkifly, Ahmad Hafiz, Sha'ban, Munirah
Format:	Conference or Workshop Item
Language:	English
Published:	2016
Subjects:	TA164 Bioengineering
Online Access:	http://irep.iium.edu.my/54993/ http://irep.iium.edu.my/54993/1/Yusof%20KRW2016%20Abstract.pdf

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recordtype	eprints
spelling	iium-549932017-02-13T06:13:33Z http://irep.iium.edu.my/54993/ Monolayer culture expansion of Annulus fibrosus cells for intervertebral disc regeneration Mohamad, Mohd Yusof Mohamed Amin, Muhammad Azri Ifwat Zulkifly, Ahmad Hafiz Sha'ban, Munirah TA164 Bioengineering Objectives/Research Problem: Tissue engineering (TE) provides an alternative approach in regenerating human tissues for clinical transplantation and drugs development purposes. In principle, TE requires good cell source, compatible biomaterial scaffold and proper signalling factors. Regeneration of intervertebral disc (IVD) structure may be achieved by establishing well-balanced between the above three TE principles. Annulus fibrosus (AF) is a potential cell source for fabrication of IVD-engineered tissue. Anatomically, AF surrounds the inner nucleus pulposus region and forms the outer layer of IVD. Being one of the important IVD structures, much attention has been given to regenerate AF. Expansion of AF cells in monolayer culture is a vital step towards cellular incorporation onto 3D biomaterial scaffold. The present study attempted to evaluate monolayer culture expansion of AF cells at different passages. Materials and Method: The rabbit’s AF cells were harvested aseptically according to the IIUM approval (IIUM/IACUCApproval/2015/[5][23]). The cells were cultured from passage 0 (P0) until passage 3 (P3). At each passage, evaluation was performed in terms of cells morphology, viability, growth kinetics, sulphated glycosaminoglycan content, DNA content, standard cytology staining and immunocytochemistry. Results and Discussion: The AF cells appeared spindle-shaped and adopted more fibroblastic traits in culture. At passage 1 (P1), AF demonstrated higher growth rate, lesser cell doubling time and significant sulphated glycosaminoglycan content when compared to other passages. No significant differences were observed in DNA content for all passages. Standard cellular staining and immunocytochemistry findings confirm the presence of pericellular matrix and cartilage specific marker i.e. collagen type II respectively. Presence of collagen type I was also noted, indicative of AF’s fibrocartilaginous nature. Conclusion: Current findings suggest expansion of AF at P1 may become a potential cell source for certain application in tissue engineering. Future improvement may be made by adding growth factors to promote the proliferation of AF cells. 2016-11-21 Conference or Workshop Item PeerReviewed application/pdf en http://irep.iium.edu.my/54993/1/Yusof%20KRW2016%20Abstract.pdf Mohamad, Mohd Yusof and Mohamed Amin, Muhammad Azri Ifwat and Zulkifly, Ahmad Hafiz and Sha'ban, Munirah (2016) Monolayer culture expansion of Annulus fibrosus cells for intervertebral disc regeneration. In: KAHS Research Week (KRW) 2016 in conjunction with 1st Allied Health Scientific Colloquium (AHSC) 2016, 21 - 25 November 2016, KAHS Examination Hall & Lecture Hall 1.
repository_type	Digital Repository
institution_category	Local University
institution	International Islamic University Malaysia
building	IIUM Repository
collection	Online Access
language	English
topic	TA164 Bioengineering
spellingShingle	TA164 Bioengineering Mohamad, Mohd Yusof Mohamed Amin, Muhammad Azri Ifwat Zulkifly, Ahmad Hafiz Sha'ban, Munirah Monolayer culture expansion of Annulus fibrosus cells for intervertebral disc regeneration
description	Objectives/Research Problem: Tissue engineering (TE) provides an alternative approach in regenerating human tissues for clinical transplantation and drugs development purposes. In principle, TE requires good cell source, compatible biomaterial scaffold and proper signalling factors. Regeneration of intervertebral disc (IVD) structure may be achieved by establishing well-balanced between the above three TE principles. Annulus fibrosus (AF) is a potential cell source for fabrication of IVD-engineered tissue. Anatomically, AF surrounds the inner nucleus pulposus region and forms the outer layer of IVD. Being one of the important IVD structures, much attention has been given to regenerate AF. Expansion of AF cells in monolayer culture is a vital step towards cellular incorporation onto 3D biomaterial scaffold. The present study attempted to evaluate monolayer culture expansion of AF cells at different passages. Materials and Method: The rabbit’s AF cells were harvested aseptically according to the IIUM approval (IIUM/IACUCApproval/2015/[5][23]). The cells were cultured from passage 0 (P0) until passage 3 (P3). At each passage, evaluation was performed in terms of cells morphology, viability, growth kinetics, sulphated glycosaminoglycan content, DNA content, standard cytology staining and immunocytochemistry. Results and Discussion: The AF cells appeared spindle-shaped and adopted more fibroblastic traits in culture. At passage 1 (P1), AF demonstrated higher growth rate, lesser cell doubling time and significant sulphated glycosaminoglycan content when compared to other passages. No significant differences were observed in DNA content for all passages. Standard cellular staining and immunocytochemistry findings confirm the presence of pericellular matrix and cartilage specific marker i.e. collagen type II respectively. Presence of collagen type I was also noted, indicative of AF’s fibrocartilaginous nature. Conclusion: Current findings suggest expansion of AF at P1 may become a potential cell source for certain application in tissue engineering. Future improvement may be made by adding growth factors to promote the proliferation of AF cells.
format	Conference or Workshop Item
author	Mohamad, Mohd Yusof Mohamed Amin, Muhammad Azri Ifwat Zulkifly, Ahmad Hafiz Sha'ban, Munirah
author_facet	Mohamad, Mohd Yusof Mohamed Amin, Muhammad Azri Ifwat Zulkifly, Ahmad Hafiz Sha'ban, Munirah
author_sort	Mohamad, Mohd Yusof
title	Monolayer culture expansion of Annulus fibrosus cells for intervertebral disc regeneration
title_short	Monolayer culture expansion of Annulus fibrosus cells for intervertebral disc regeneration
title_full	Monolayer culture expansion of Annulus fibrosus cells for intervertebral disc regeneration
title_fullStr	Monolayer culture expansion of Annulus fibrosus cells for intervertebral disc regeneration
title_full_unstemmed	Monolayer culture expansion of Annulus fibrosus cells for intervertebral disc regeneration
title_sort	monolayer culture expansion of annulus fibrosus cells for intervertebral disc regeneration
publishDate	2016
url	http://irep.iium.edu.my/54993/ http://irep.iium.edu.my/54993/1/Yusof%20KRW2016%20Abstract.pdf
first_indexed	2023-09-18T21:17:45Z
last_indexed	2023-09-18T21:17:45Z
_version_	1777411675945697280

Monolayer culture expansion of Annulus fibrosus cells for intervertebral disc regeneration

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