Expression analysis of Notch signaling pathway molecules in SHED cultured in keratinocyte growth medium

Authors:
Siti Aisyah Mohd Taha, Joanne Koh Su Ling, Nur Izyan Binti Azmi, Thirumulu Ponnuraj Kannan, Ahmad Azlina, Khairani Idah Mokhtar
In:
Source: Braz J Oral Sci
Publication Date: (2015)
Issue: 14(2): 135-140
Research Area:
Stem Cells
Basic Research
Cells used in publication:
DPSC (dental pulp stem cells)
Species: human
Tissue Origin: teeth
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.