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

• Main Authors: Abdullah, N.A (Author), Berahim, Z. (Author), Farea, M. (Author), Halim, A.S (Author), Husein, A. (Author), Lim, C.K (Author), Mokhtar, K. (Author), Mokhtar, K.I (Author), Rani, A.Q (Author)
• Format: Article
• Language: English
• Published: MDPI AG 2013
• Subjects: adult; amelogenin; Amelogenin; article; cadherin; Cadherins; CD34 antigen; cell culture; cell isolation; cell proliferation; Cell Proliferation; cell separation; Cell Separation; cell shape; Cells, Cultured; Characterization; controlled study; cytokeratin; cytokeratin 14; cytokeratin 14 gene; cytology; edetic acid; Enamel Organ; endoglin; epithelium cell; fibroblast; flow cytometry; Flow Cytometry; fluorescent antibody technique; Fluorescent Antibody Technique; gene expression regulation; Gene Expression Regulation; genetics; glyceraldehyde 3 phosphate dehydrogenase; glyceraldehyde 3 phosphate dehydrogenase gene; Hermes antigen; Hertwig epithelial root sheat cell; Hertwig's epithelial root sheath (HERS); Homogenous cell culture; human; human cell; Humans; immunofluorescence test; in vitro study; keratin; keratinocyte; Keratinocytes; Keratins; metabolism; p63 gene; procedures; protein p63; reverse transcription polymerase chain reaction; Selective digestion; tooth flora; tooth root; Tooth Root; TP63 protein, human; transcription factor; Transcription Factors; trypsin; tumor suppressor protein; Tumor Suppressor Proteins; uvomorulin; uvomorulin gene
• Online Access: View Fulltext in Publisher; View in Scopus
• ISBN: 16616596 (ISSN)
• DOI: 10.3390/ijms140611157

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.