title: Isolation and enhancement of a homogenous in vitro human Hertwig's epithelial root sheath cell population.
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.
authors.affiliation: Conservative Department, School of Dental Sciences, Universiti Sains Malaysia, 16150 Kubang Kerian, Kelantan, Malaysia. ghorafi44@yahoo.com.
authors.forename: Manal
authors.initials: M
authors.lastname: Farea
authors.forename: Ahmad Sukari
authors.initials: AS
authors.lastname: Halim
authors.forename: Nurul Asma
authors.initials: NA
authors.lastname: Abdullah
authors.forename: Chin Keong
authors.initials: CK
authors.lastname: Lim
authors.forename: Khairani Idah
authors.initials: KI
authors.lastname: Mokhtar
authors.forename: Zurairah
authors.initials: Z
authors.lastname: Berahim
authors.forename: Kasmawati
authors.initials: K
authors.lastname: Mokhtar
authors.forename: Abdul Qawee
authors.initials: AQ
authors.lastname: Rani
authors.forename: Adam
authors.initials: A
authors.lastname: Husein
mesh_terms: D053523:Amelogenin / Q000235:genetics / Q000378:metabolism; D015820:Cadherins / Q000235:genetics / Q000378:metabolism; D049109:Cell Proliferation; D002469:Cell Separation / Q000379:methods*; D002478:Cells, Cultured; D004658:Enamel Organ / Q000166:cytology*; D005434:Flow Cytometry; D005455:Fluorescent Antibody Technique; D005786:Gene Expression Regulation; D006801:Humans; D015603:Keratinocytes / Q000166:cytology / Q000378:metabolism; D007633:Keratins / Q000235:genetics / Q000378:metabolism; D014092:Tooth Root / Q000166:cytology*; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism; D025521:Tumor Suppressor Proteins / Q000235:genetics / Q000378:metabolism
chemical_list: D053523:Amelogenin; D015820:Cadherins; C115051:TP63 protein, human; D014157:Transcription Factors; D025521:Tumor Suppressor Proteins; D007633:Keratins