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)
Abstract
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.
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