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

International Journal of Molecular Sciences

Manal Farea²  Ahmad Sukari Halim³  Nurul Asma Abdullah⁴  Chin Keong Lim¹  Khairani Idah Mokhtar⁴  Zurairah Berahim²  Kasmawati Mokhtar²  Abdul Qawee Rani⁵ 

[1] Centre of Studies for Preclinical Science, Faculty of Dentistry, Universiti Teknologi MARA, 40450 Shah Alam, Selangor, Malaysia; E-Mail:;Conservative Department, School of Dental Sciences, Universiti Sains Malaysia, 16150 Kubang Kerian, Kelantan, Malaysia; E-Mails:;Reconstructive Sciences Unit, School of Medical Sciences, Universiti Sains Malaysia, 16150 Kubang Kerian, Kelantan, Malaysia;Oral Biology Unit, School of Dental Sciences, Universiti Sains Malaysia, 16150 Kubang Kerian, Kelantan, Malaysia; E-Mails:;Human Genome Centre, School of Medical Sciences, Universiti Sains Malaysia, 16150 Kubang Kerian, Kelantan, Malaysia; E-Mail:

关键词: Hertwig’s epithelial root sheath (HERS);    homogenous cell culture;    selective digestion;    characterization;   

DOI  :  10.3390/ijms140611157

来源: mdpi

【 摘 要 】

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.

【 授权许可】

CC BY   
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