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Expression analysis of Notch signaling pathway molecules in SHED cultured in keratinocyte growth medium
Taha, Siti Aisyah Mohd; Ling, Joanne Koh Su; Binti Azmi, Nur Izyan; Kannan, Thirumulu Ponnuraj; Azlina, Ahmad & Mokhtar, Khairani Idah
Abstract
Aim: To detect the expression of molecules associated with Notch signaling pathway in stem cells
from human exfoliated deciduous teeth (SHED) cultured in specific differentiation medium, namely,
keratinocyte growth medium (KGM). Methods: RNA was extracted from SHED harvested on
day 1, 3 and 7. RNA was reverse-transcribed to obtain the cDNA and then proceeded with PCR
using specific primers for the Notch signaling pathway molecules (Notch1, Jagged-1, Jagged-2
and, Hes1) as well as stem cell marker (Nanog). PCR products were electrophoresed on a 2%
agarose gel and stained with SYBR green. Results: Notch-1 was highly expressed in SHED
cultured in KGM and showed increase in density as the days progressed, while Jagged-1
showed a decrease. Jagged-2 on the other hand, showed a slight increase on day 3 followed
by a decrease on day 7. However, Hes-1 was not expressed in SHED cultured in KGM. Nanog
showed expression only on day 3 and gradually increased in expression on day 7.
Conclusions: Notch signaling pathway associated molecules; Notch-1, Jagged-1, Jagged-2,
and stem cell marker Nanog are expressed in SHED cultured in KGM which may be involved in
the differentiation into epithelial-like cells in human dental pulp tissues.
Keywords
receptors; notch; gene expression; stem cells; tooth; deciduous; culture media
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