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

• Main Authors: Manal Farea, Ahmad Sukari Halim, Nurul Asma Abdullah, Chin Keong Lim, Khairani Idah Mokhtar, Zurairah Berahim, Kasmawati Mokhtar, Abdul Qawee Rani, Adam Husein
• Format: Article
• Language: English
• Published: MDPI AG 2013-05-01
• Series: International Journal of Molecular Sciences
• Subjects: Hertwig’s epithelial root sheath (HERS); homogenous cell culture; selective digestion; characterization
• Online Access: http://www.mdpi.com/1422-0067/14/6/11157

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.