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

Siti Aisyah Mohd Taha,Khairani Idah Mokhtar,Nur Izyan Binti Azmi,Joanne Koh Su Ling,Ahmad Azlina,Thirumulu Ponnuraj Kannan

doi:10.1590/1677-3225v14n2a08

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.

Highlights

Received for publication: April 09, 2015 Accepted: June 09, 2015Correspondence to: Thirumulu Ponnuraj Kannan School of Dental Sciences, Universiti Sains MalaysiaBraz J Oral Sci. 14(2)[135-140]Stem cells from human exfoliated deciduous teeth (SHED) are multipotent stem cells derived from the pulp tissues of extracted deciduous teeth[1]
The samples analyzed for the expression of Notch-1 receptor in this study cultured in keratinocyte growth medium (KGM) showed expression of Notch-1, which increased in expression as the days progressed from day 1 to, 3 and 7
The DSL-type ligand, a Notch-type receptor and a transcription factor of the CSL family are the three different molecules which are the main components of Notch signaling pathway[3,12]

Summary

Introduction

Received for publication: April 09, 2015 Accepted: June 09, 2015Correspondence to: Thirumulu Ponnuraj Kannan School of Dental Sciences, Universiti Sains MalaysiaBraz J Oral Sci. 14(2)[135-140]Stem cells from human exfoliated deciduous teeth (SHED) are multipotent stem cells derived from the pulp tissues of extracted deciduous teeth[1]. SHED has the ability to be differentiated to specific cell lineages such as odontoblasts and osteoblasts as well as epithelial like cells. SHED was able to differentiate into epithelial like cells when cultured in keratinocyte growth medium (KGM)[2]. Since the Notch signaling pathway molecules play an important role in differentiation of epithelial cells, it is important to identify the presence of notch signaling molecules in SHED during the process of cell differentiation. The Notch signaling pathway provides important intercellular signaling mechanisms essential for cell fate specification and it regulates differentiation and proliferation of stem or progenitor cells by para-inducing effects[3,4]. The core components of a Notch signaling pathway involves three different molecules; the DSL-type ligand, a Notch-type receptor and a transcription factor of the CSL family

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