Isolation and enhancement of a homogenous in vitro human hertwig's epithelial root sheath cell population

• Published in: International Journal of Molecular Sciences
• Main Author: Farea M.; Halim A.S.; Abdullah N.A.; Lim C.K.; Mokhtar K.I.; Berahim Z.; Mokhtar K.; Rani A.Q.; Husein A.
• Format: Article
• Language: English
• Published: MDPI AG 2013
• Online Access: https://www.scopus.com/inward/record.uri?eid=2-s2.0-84878495678&doi=10.3390%2fijms140611157&partnerID=40&md5=6a8c8c648d9bc959c113254b70f56456

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. © 2013 by the authors; licensee MDPI, Basel, Switzerland.