TGFβ-1 and ALK5 inhibitor treatment affects expressions of TGF β signalling pathway associated molecules in SHED cultured in keratinocyte growth medium

TGFβ-1 and ALK5 inhibitor treatment affects expressions of TGF β signalling pathway associated molecules in SHED cultured in keratinocyte growth medium

Stem cells from human exfoliated deciduous teeth (SHED) can be induced to differentiate into epithelial-like cells when cultured in a differentiation medium. Transforming growth factor beta (TGF-β) signalling pathway plays an important regulatory function in epithelial proliferation and differentiat...

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Main Authors: Mokhtar@Makhtar, Khairani Idah, Azmi, Nur Izyan, Berahim, Zurairah, Kannan, Thirumulu Ponnuraj, Ahmad, Azlina
Format: Conference or Workshop Item
Language:English
Published: 2015
Subjects:
RK Dentistry
Online Access:http://irep.iium.edu.my/44828/
http://irep.iium.edu.my/44828/1/44828.pdf
id iium-44828
recordtype eprints
spelling iium-448282015-10-05T06:42:53Z http://irep.iium.edu.my/44828/ TGFβ-1 and ALK5 inhibitor treatment affects expressions of TGF β signalling pathway associated molecules in SHED cultured in keratinocyte growth medium Mokhtar@Makhtar, Khairani Idah Azmi, Nur Izyan Berahim, Zurairah Kannan, Thirumulu Ponnuraj Ahmad, Azlina RK Dentistry Stem cells from human exfoliated deciduous teeth (SHED) can be induced to differentiate into epithelial-like cells when cultured in a differentiation medium. Transforming growth factor beta (TGF-β) signalling pathway plays an important regulatory function in epithelial proliferation and differentiation.TGF-β1 functions through a complex mechanism consisting of TGFβ type II and type I (ALK5) receptor to initiate cellular processes which could be inhibited by SB431542 molecule (ALK5 inhibitor). This study aims to analyse the expression of TGF-β signalling pathway associated molecules, ALK5 and Smad4, in SHED cultured in keratinocyte growth medium (KGM), treated with exogenous TGF-β1 and ALK5 inhibitor. SHED was cultured in KGM treated with TGF-β1 or ALK-5 inhibitor. RNA was extracted on cells harvested on day 1, 3, 7, 14 and 21 and subjected to reverse transcriptase-PCR using specific primers for ALK5, Smad4 and stem cell marker (Nanog). PCR products were electrophoresed on a 2% agarose gel stained with SYBR green. Nanog expression was maintained from day1 to 14 in all samples but was down regulated at day 21. ALK5 was expressed in most of the samples analysed from day 1 to 14 and down regulated at day 21. Smad4 expression was up regulated at day 1 to 3, down regulated from day 7 to 14 and lost its expression at day 21. TGFβ-1 and ALK5 inhibitor treatment affects the expressions of Nanog, ALK5 and Smad4 in SHED cultured in KGM. The involvement of these molecules during the differentiation process of SHED into epithelial-like cells will contribute further in the fundamental aspects of tissue regeneration research and highlights the importance of dental pulp stem cell technology. 2015-09-08 Conference or Workshop Item PeerReviewed application/pdf en http://irep.iium.edu.my/44828/1/44828.pdf Mokhtar@Makhtar, Khairani Idah and Azmi, Nur Izyan and Berahim, Zurairah and Kannan, Thirumulu Ponnuraj and Ahmad, Azlina (2015) TGFβ-1 and ALK5 inhibitor treatment affects expressions of TGF β signalling pathway associated molecules in SHED cultured in keratinocyte growth medium. In: 22nd Annual Scientific Meeting of Malaysian Society for Molecular Biology and Biotechnology, 8th-9th September 2015, RMIC, University of Malaya.
repository_type Digital Repository
institution_category Local University
institution International Islamic University Malaysia
building IIUM Repository
collection Online Access
language English
topic RK Dentistry
spellingShingle RK Dentistry
Mokhtar@Makhtar, Khairani Idah
Azmi, Nur Izyan
Berahim, Zurairah
Kannan, Thirumulu Ponnuraj
Ahmad, Azlina
TGFβ-1 and ALK5 inhibitor treatment affects expressions of TGF β signalling pathway associated molecules in SHED cultured in keratinocyte growth medium
description Stem cells from human exfoliated deciduous teeth (SHED) can be induced to differentiate into epithelial-like cells when cultured in a differentiation medium. Transforming growth factor beta (TGF-β) signalling pathway plays an important regulatory function in epithelial proliferation and differentiation.TGF-β1 functions through a complex mechanism consisting of TGFβ type II and type I (ALK5) receptor to initiate cellular processes which could be inhibited by SB431542 molecule (ALK5 inhibitor). This study aims to analyse the expression of TGF-β signalling pathway associated molecules, ALK5 and Smad4, in SHED cultured in keratinocyte growth medium (KGM), treated with exogenous TGF-β1 and ALK5 inhibitor. SHED was cultured in KGM treated with TGF-β1 or ALK-5 inhibitor. RNA was extracted on cells harvested on day 1, 3, 7, 14 and 21 and subjected to reverse transcriptase-PCR using specific primers for ALK5, Smad4 and stem cell marker (Nanog). PCR products were electrophoresed on a 2% agarose gel stained with SYBR green. Nanog expression was maintained from day1 to 14 in all samples but was down regulated at day 21. ALK5 was expressed in most of the samples analysed from day 1 to 14 and down regulated at day 21. Smad4 expression was up regulated at day 1 to 3, down regulated from day 7 to 14 and lost its expression at day 21. TGFβ-1 and ALK5 inhibitor treatment affects the expressions of Nanog, ALK5 and Smad4 in SHED cultured in KGM. The involvement of these molecules during the differentiation process of SHED into epithelial-like cells will contribute further in the fundamental aspects of tissue regeneration research and highlights the importance of dental pulp stem cell technology.
format Conference or Workshop Item
author Mokhtar@Makhtar, Khairani Idah
Azmi, Nur Izyan
Berahim, Zurairah
Kannan, Thirumulu Ponnuraj
Ahmad, Azlina
author_facet Mokhtar@Makhtar, Khairani Idah
Azmi, Nur Izyan
Berahim, Zurairah
Kannan, Thirumulu Ponnuraj
Ahmad, Azlina
author_sort Mokhtar@Makhtar, Khairani Idah
title TGFβ-1 and ALK5 inhibitor treatment affects expressions of TGF β signalling pathway associated molecules in SHED cultured in keratinocyte growth medium
title_short TGFβ-1 and ALK5 inhibitor treatment affects expressions of TGF β signalling pathway associated molecules in SHED cultured in keratinocyte growth medium
title_full TGFβ-1 and ALK5 inhibitor treatment affects expressions of TGF β signalling pathway associated molecules in SHED cultured in keratinocyte growth medium
title_fullStr TGFβ-1 and ALK5 inhibitor treatment affects expressions of TGF β signalling pathway associated molecules in SHED cultured in keratinocyte growth medium
title_full_unstemmed TGFβ-1 and ALK5 inhibitor treatment affects expressions of TGF β signalling pathway associated molecules in SHED cultured in keratinocyte growth medium
title_sort tgfβ-1 and alk5 inhibitor treatment affects expressions of tgf β signalling pathway associated molecules in shed cultured in keratinocyte growth medium
publishDate 2015
url http://irep.iium.edu.my/44828/
http://irep.iium.edu.my/44828/1/44828.pdf
first_indexed 2023-09-18T21:03:44Z
last_indexed 2023-09-18T21:03:44Z
_version_ 1777410795027562496

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