An In Vitro Barrier Model of the Human Submandibular Salivary Gland Epithelium Based on a Single Cell Clone of Cell Line HTB-41: Establishment and Application for Biomarker Transport Studies
The blood–saliva barrier (BSB) consists of the sum of the epithelial cell layers of the oral mucosa and salivary glands. In vitro models of the BSB are inevitable to investigate and understand the transport of salivary biomarkers from blood to saliva. Up to now, standardized, cell line-based models...
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MDPI AG
2020-08-01
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| Series: | Biomedicines |
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| Online Access: | https://www.mdpi.com/2227-9059/8/9/302 |
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| author | Grace C. Lin Merima Smajlhodzic Anna-Maria Bandian Heinz-Peter Friedl Tamara Leitgeb Sabrina Oerter Kerstin Stadler Ulrich Giese Johannes R. Peham Lynne Bingle Winfried Neuhaus |
| author_facet | Grace C. Lin Merima Smajlhodzic Anna-Maria Bandian Heinz-Peter Friedl Tamara Leitgeb Sabrina Oerter Kerstin Stadler Ulrich Giese Johannes R. Peham Lynne Bingle Winfried Neuhaus |
| author_sort | Grace C. Lin |
| collection | DOAJ |
| description | The blood–saliva barrier (BSB) consists of the sum of the epithelial cell layers of the oral mucosa and salivary glands. In vitro models of the BSB are inevitable to investigate and understand the transport of salivary biomarkers from blood to saliva. Up to now, standardized, cell line-based models of the epithelium of the submandibular salivary gland are still missing for this purpose. Therefore, we established epithelial barrier models of the submandibular gland derived from human cell line HTB-41 (A-253). Single clone isolation resulted in five different clones (B2, B4, B9, D3, and F11). Clones were compared to the parental cell line HTB-41 using measurements of the transepithelial electrical resistance (TEER), paracellular marker permeability assays and analysis of marker expression for acinar, ductal, and myoepithelial cells. Two clones (B9, D3) were characterized to be of acinar origin, one clone (F11) to be of myoepithelial origin and one isolation (B4) derived from two cells, to be presumably a mixture of acinar and ductal origin. Clone B2, presumably of ductal origin, showed a significantly higher paracellular barrier compared to other clones and parental HTB-41. The distinct molecular identity of clone B2 was confirmed by immunofluorescent staining, qPCR, and flow cytometry. Experiments with ferritin, a biomarker for iron storage, demonstrated the applicability of the selected model based on clone B2 for transport studies. In conclusion, five different clones originating from the submandibular gland cell line HTB-41 were successfully characterized and established as epithelial barrier models. Studies with the model based on the tightest clone B2 confirmed its suitability for transport studies in biomarker research. |
| first_indexed | 2024-03-10T16:57:42Z |
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| institution | Directory Open Access Journal |
| issn | 2227-9059 |
| language | English |
| last_indexed | 2024-03-10T16:57:42Z |
| publishDate | 2020-08-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Biomedicines |
| spelling | doaj.art-32aead8fe94e4ab09566257631bd2d892023-11-20T11:03:47ZengMDPI AGBiomedicines2227-90592020-08-018930210.3390/biomedicines8090302An In Vitro Barrier Model of the Human Submandibular Salivary Gland Epithelium Based on a Single Cell Clone of Cell Line HTB-41: Establishment and Application for Biomarker Transport StudiesGrace C. Lin0Merima Smajlhodzic1Anna-Maria Bandian2Heinz-Peter Friedl3Tamara Leitgeb4Sabrina Oerter5Kerstin Stadler6Ulrich Giese7Johannes R. Peham8Lynne Bingle9Winfried Neuhaus10Competence Unit Molecular Diagnostics, Center for Health and Bioresources, Austrian Institute of Technology (AIT) GmbH, Giefinggasse 4, 1210 Vienna, AustriaCompetence Unit Molecular Diagnostics, Center for Health and Bioresources, Austrian Institute of Technology (AIT) GmbH, Giefinggasse 4, 1210 Vienna, AustriaCompetence Unit Molecular Diagnostics, Center for Health and Bioresources, Austrian Institute of Technology (AIT) GmbH, Giefinggasse 4, 1210 Vienna, AustriaCompetence Unit Molecular Diagnostics, Center for Health and Bioresources, Austrian Institute of Technology (AIT) GmbH, Giefinggasse 4, 1210 Vienna, AustriaCompetence Unit Molecular Diagnostics, Center for Health and Bioresources, Austrian Institute of Technology (AIT) GmbH, Giefinggasse 4, 1210 Vienna, AustriaFraunhofer Institute for Silicate Research (ISC), Neunerplatz 2, 97082 Würzburg, GermanyCompetence Unit Molecular Diagnostics, Center for Health and Bioresources, Austrian Institute of Technology (AIT) GmbH, Giefinggasse 4, 1210 Vienna, AustriaCompetence Unit Molecular Diagnostics, Center for Health and Bioresources, Austrian Institute of Technology (AIT) GmbH, Giefinggasse 4, 1210 Vienna, AustriaCompetence Unit Molecular Diagnostics, Center for Health and Bioresources, Austrian Institute of Technology (AIT) GmbH, Giefinggasse 4, 1210 Vienna, AustriaSchool of Clinical Dentistry, University of Sheffield, Claremont Crescent, Sheffield S10 2TA, UKCompetence Unit Molecular Diagnostics, Center for Health and Bioresources, Austrian Institute of Technology (AIT) GmbH, Giefinggasse 4, 1210 Vienna, AustriaThe blood–saliva barrier (BSB) consists of the sum of the epithelial cell layers of the oral mucosa and salivary glands. In vitro models of the BSB are inevitable to investigate and understand the transport of salivary biomarkers from blood to saliva. Up to now, standardized, cell line-based models of the epithelium of the submandibular salivary gland are still missing for this purpose. Therefore, we established epithelial barrier models of the submandibular gland derived from human cell line HTB-41 (A-253). Single clone isolation resulted in five different clones (B2, B4, B9, D3, and F11). Clones were compared to the parental cell line HTB-41 using measurements of the transepithelial electrical resistance (TEER), paracellular marker permeability assays and analysis of marker expression for acinar, ductal, and myoepithelial cells. Two clones (B9, D3) were characterized to be of acinar origin, one clone (F11) to be of myoepithelial origin and one isolation (B4) derived from two cells, to be presumably a mixture of acinar and ductal origin. Clone B2, presumably of ductal origin, showed a significantly higher paracellular barrier compared to other clones and parental HTB-41. The distinct molecular identity of clone B2 was confirmed by immunofluorescent staining, qPCR, and flow cytometry. Experiments with ferritin, a biomarker for iron storage, demonstrated the applicability of the selected model based on clone B2 for transport studies. In conclusion, five different clones originating from the submandibular gland cell line HTB-41 were successfully characterized and established as epithelial barrier models. Studies with the model based on the tightest clone B2 confirmed its suitability for transport studies in biomarker research.https://www.mdpi.com/2227-9059/8/9/302salivary glandsubmandibularblood–saliva barrierSjögren’s syndromerheumatoid arthritisperiodontitis |
| spellingShingle | Grace C. Lin Merima Smajlhodzic Anna-Maria Bandian Heinz-Peter Friedl Tamara Leitgeb Sabrina Oerter Kerstin Stadler Ulrich Giese Johannes R. Peham Lynne Bingle Winfried Neuhaus An In Vitro Barrier Model of the Human Submandibular Salivary Gland Epithelium Based on a Single Cell Clone of Cell Line HTB-41: Establishment and Application for Biomarker Transport Studies Biomedicines salivary gland submandibular blood–saliva barrier Sjögren’s syndrome rheumatoid arthritis periodontitis |
| title | An In Vitro Barrier Model of the Human Submandibular Salivary Gland Epithelium Based on a Single Cell Clone of Cell Line HTB-41: Establishment and Application for Biomarker Transport Studies |
| title_full | An In Vitro Barrier Model of the Human Submandibular Salivary Gland Epithelium Based on a Single Cell Clone of Cell Line HTB-41: Establishment and Application for Biomarker Transport Studies |
| title_fullStr | An In Vitro Barrier Model of the Human Submandibular Salivary Gland Epithelium Based on a Single Cell Clone of Cell Line HTB-41: Establishment and Application for Biomarker Transport Studies |
| title_full_unstemmed | An In Vitro Barrier Model of the Human Submandibular Salivary Gland Epithelium Based on a Single Cell Clone of Cell Line HTB-41: Establishment and Application for Biomarker Transport Studies |
| title_short | An In Vitro Barrier Model of the Human Submandibular Salivary Gland Epithelium Based on a Single Cell Clone of Cell Line HTB-41: Establishment and Application for Biomarker Transport Studies |
| title_sort | in vitro barrier model of the human submandibular salivary gland epithelium based on a single cell clone of cell line htb 41 establishment and application for biomarker transport studies |
| topic | salivary gland submandibular blood–saliva barrier Sjögren’s syndrome rheumatoid arthritis periodontitis |
| url | https://www.mdpi.com/2227-9059/8/9/302 |
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