Int. J. Mol. Sci. 2013, 14(6), 11157-11170; doi:10.3390/ijms140611157
Article

Isolation and Enhancement of a Homogenous in Vitro Human Hertwig’s Epithelial Root Sheath Cell Population

1 Conservative Department, School of Dental Sciences, Universiti Sains Malaysia, 16150 Kubang Kerian, Kelantan, Malaysia 2 Reconstructive Sciences Unit, School of Medical Sciences, Universiti Sains Malaysia, 16150 Kubang Kerian, Kelantan, Malaysia 3 Oral Biology Unit, School of Dental Sciences, Universiti Sains Malaysia, 16150 Kubang Kerian, Kelantan, Malaysia 4 Centre of Studies for Preclinical Science, Faculty of Dentistry, Universiti Teknologi MARA, 40450 Shah Alam, Selangor, Malaysia 5 Human Genome Centre, School of Medical Sciences, Universiti Sains Malaysia, 16150 Kubang Kerian, Kelantan, Malaysia
* Authors to whom correspondence should be addressed.
Received: 9 April 2013; in revised form: 9 May 2013 / Accepted: 22 May 2013 / Published: 27 May 2013
(This article belongs to the Section Biochemistry, Molecular Biology and Biophysics)
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Abstract: Hertwig’s epithelial root sheath (HERS) cells play a pivotal role during root formation of the tooth and are able to form cementum-like tissue. The aim of the present study was to establish a HERS cell line for molecular and biochemical studies using a selective digestion method. Selective digestion was performed by the application of trypsin-EDTA for 2 min, which led to the detachment of fibroblast-like-cells, with the rounded cells attached to the culture plate. The HERS cells displayed a typical cuboidal/squamous-shaped appearance. Characterization of the HERS cells using immunofluorescence staining and flow cytometry analysis showed that these cells expressed pan-cytokeratin, E-cadherin, and p63 as epithelial markers. Moreover, RT-PCR confirmed that these cells expressed epithelial-related genes, such as cytokeratin 14, E-cadherin, and ΔNp63. Additionally, HERS cells showed low expression of CD44 and CD105 with absence of CD34 and amelogenin expressions. In conclusion, HERS cells have been successfully isolated using a selective digestion method, thus enabling future studies on the roles of these cells in the formation of cementum-like tissue in vitro.
Keywords: Hertwig’s epithelial root sheath (HERS); homogenous cell culture; selective digestion; characterization

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MDPI and ACS Style

Farea, M.; Halim, A.S.; Abdullah, N.A.; Lim, C.K.; Mokhtar, K.I.; Berahim, Z.; Mokhtar, K.; Rani, A.Q.; Husein, A. Isolation and Enhancement of a Homogenous in Vitro Human Hertwig’s Epithelial Root Sheath Cell Population. Int. J. Mol. Sci. 2013, 14, 11157-11170.

AMA Style

Farea M, Halim AS, Abdullah NA, Lim CK, Mokhtar KI, Berahim Z, Mokhtar K, Rani AQ, Husein A. Isolation and Enhancement of a Homogenous in Vitro Human Hertwig’s Epithelial Root Sheath Cell Population. International Journal of Molecular Sciences. 2013; 14(6):11157-11170.

Chicago/Turabian Style

Farea, Manal; Halim, Ahmad S.; Abdullah, Nurul A.; Lim, Chin K.; Mokhtar, Khairani I.; Berahim, Zurairah; Mokhtar, Kasmawati; Rani, Abdul Q.; Husein, Adam. 2013. "Isolation and Enhancement of a Homogenous in Vitro Human Hertwig’s Epithelial Root Sheath Cell Population." Int. J. Mol. Sci. 14, no. 6: 11157-11170.

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