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Role of Transforming Growth Factor-Beta 1 (TGFB-1) and the cell signaling pathway in the differentiation of SHED into epithelial-like cells

Epithelial cells plays crucial role in human body and in oral cavity, epithelial formation is important during the healing process after injuries. Previous study had shown that primary cells from human exfoliated deciduous teeth (SHED) can be differentiated into epithelial-like cells when culture...

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Main Author: Ahmad, Azlina
Format: Monograph
Language:English
Published: Universiti Sains Malaysia 2017
Subjects:
QR Microbiology
Online Access:http://eprints.usm.my/57036/1/DR%20AZLINA%20AHMAD-Eprints.pdf
http://eprints.usm.my/57036/
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spelling my.usm.eprints.57036 http://eprints.usm.my/57036/ Role of Transforming Growth Factor-Beta 1 (TGFB-1) and the cell signaling pathway in the differentiation of SHED into epithelial-like cells Ahmad, Azlina QR Microbiology Epithelial cells plays crucial role in human body and in oral cavity, epithelial formation is important during the healing process after injuries. Previous study had shown that primary cells from human exfoliated deciduous teeth (SHED) can be differentiated into epithelial-like cells when cultured in Keratinocyte Growth Medium (K.GM). This study aim to determine the effects of TGF-p 1 and ALK-5 inhibitor in the differentiation of SHED into epithelial-like cells cultured in specific differentiation medium (KGM). MTT and alamarBlue assay was done to investigate and determine the best concentration to be used in the study. Then, gene expression of mesenchymal, epithelial, and TGF-P signaling marker was carried out on SHED cultured with KGM treated with 1.25 ng/ml of TGF-PI or 0.625 pM of ALK-5 inhibitor. The treated SHED was harvested at day 1,3,7,14, and 21 and subjected to RNA extraction. Then, study was proceeded with two-step Reverse Transcriptase Polymerase Chain Reaction (RT-PCR). The presence of mesenchymal marker, Nanog on all samples except on SHED cultured in KGM + 0.625 pM on day 21. Based on epithelial marker, Keratin5 showed there is no expression on all samples treatment except control which is keratinocyte cells. For TGF-p signaling marker which were ALK-5 and Srnad4, Smad4 showed expression on day 1 until 7 for all treatmets and only SPIED in KGM on day 14 showed a mild expression. ALK5 expressed in most of samples analysed but only SHED in KGM with TGF-P 1 showed no expression. As conclusion, this study showed that TGF-P I and ALK-5 inhibitor might give a little effects to the differentiation of SHED into epithelial-like cells but not enough to induce the differentiation process. Universiti Sains Malaysia 2017 Monograph NonPeerReviewed application/pdf en http://eprints.usm.my/57036/1/DR%20AZLINA%20AHMAD-Eprints.pdf Ahmad, Azlina (2017) Role of Transforming Growth Factor-Beta 1 (TGFB-1) and the cell signaling pathway in the differentiation of SHED into epithelial-like cells. Project Report. Universiti Sains Malaysia. (Submitted)
institution Universiti Sains Malaysia
building Hamzah Sendut Library
collection Institutional Repository
continent Asia
country Malaysia
content_provider Universiti Sains Malaysia
content_source USM Institutional Repository
url_provider http://eprints.usm.my/
language English
topic QR Microbiology
spellingShingle QR Microbiology
Ahmad, Azlina
Role of Transforming Growth Factor-Beta 1 (TGFB-1) and the cell signaling pathway in the differentiation of SHED into epithelial-like cells
description Epithelial cells plays crucial role in human body and in oral cavity, epithelial formation is important during the healing process after injuries. Previous study had shown that primary cells from human exfoliated deciduous teeth (SHED) can be differentiated into epithelial-like cells when cultured in Keratinocyte Growth Medium (K.GM). This study aim to determine the effects of TGF-p 1 and ALK-5 inhibitor in the differentiation of SHED into epithelial-like cells cultured in specific differentiation medium (KGM). MTT and alamarBlue assay was done to investigate and determine the best concentration to be used in the study. Then, gene expression of mesenchymal, epithelial, and TGF-P signaling marker was carried out on SHED cultured with KGM treated with 1.25 ng/ml of TGF-PI or 0.625 pM of ALK-5 inhibitor. The treated SHED was harvested at day 1,3,7,14, and 21 and subjected to RNA extraction. Then, study was proceeded with two-step Reverse Transcriptase Polymerase Chain Reaction (RT-PCR). The presence of mesenchymal marker, Nanog on all samples except on SHED cultured in KGM + 0.625 pM on day 21. Based on epithelial marker, Keratin5 showed there is no expression on all samples treatment except control which is keratinocyte cells. For TGF-p signaling marker which were ALK-5 and Srnad4, Smad4 showed expression on day 1 until 7 for all treatmets and only SPIED in KGM on day 14 showed a mild expression. ALK5 expressed in most of samples analysed but only SHED in KGM with TGF-P 1 showed no expression. As conclusion, this study showed that TGF-P I and ALK-5 inhibitor might give a little effects to the differentiation of SHED into epithelial-like cells but not enough to induce the differentiation process.
format Monograph
author Ahmad, Azlina
author_facet Ahmad, Azlina
author_sort Ahmad, Azlina
title Role of Transforming Growth Factor-Beta 1 (TGFB-1) and the cell signaling pathway in the differentiation of SHED into epithelial-like cells
title_short Role of Transforming Growth Factor-Beta 1 (TGFB-1) and the cell signaling pathway in the differentiation of SHED into epithelial-like cells
title_full Role of Transforming Growth Factor-Beta 1 (TGFB-1) and the cell signaling pathway in the differentiation of SHED into epithelial-like cells
title_fullStr Role of Transforming Growth Factor-Beta 1 (TGFB-1) and the cell signaling pathway in the differentiation of SHED into epithelial-like cells
title_full_unstemmed Role of Transforming Growth Factor-Beta 1 (TGFB-1) and the cell signaling pathway in the differentiation of SHED into epithelial-like cells
title_sort role of transforming growth factor-beta 1 (tgfb-1) and the cell signaling pathway in the differentiation of shed into epithelial-like cells
publisher Universiti Sains Malaysia
publishDate 2017
url http://eprints.usm.my/57036/1/DR%20AZLINA%20AHMAD-Eprints.pdf
http://eprints.usm.my/57036/
_version_ 1768007871094587392
score 13.211869

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