| Autor: |
Kim J; Department of Oto-Rhino-Laryngology-Head and Neck Surgery, St.-Josefs-Hospital Hagen, University of Witten/Herdecke, D-58097 Hagen, Germany., Hegener K; Department of Oto-Rhino-Laryngology-Head and Neck Surgery, St.-Josefs-Hospital Hagen, University of Witten/Herdecke, D-58097 Hagen, Germany., Hagedorn C; Chair of Biochemistry and Molecular Medicine, Center for Biomedical Education and Research, School of Life Sciences (ZBAF), University of Witten/Herdecke, D-58453 Witten, Germany., Jamal Jameel K; Medical Clinic III for Pneumology, Allergology and Sleep Medicine, Bergmannsheil University Hospital, Ruhr-University Bochum, D-44789 Bochum, Germany., Weidinger D; Medical Clinic III for Pneumology, Allergology and Sleep Medicine, Bergmannsheil University Hospital, Ruhr-University Bochum, D-44789 Bochum, Germany., Seuthe IMC; Department of Oto-Rhino-Laryngology-Head and Neck Surgery, St.-Josefs-Hospital Hagen, University of Witten/Herdecke, D-58097 Hagen, Germany., Eichhorn S; Department of Oto-Rhino-Laryngology-Head and Neck Surgery, St.-Josefs-Hospital Hagen, University of Witten/Herdecke, D-58097 Hagen, Germany., Kreppel F; Chair of Biochemistry and Molecular Medicine, Center for Biomedical Education and Research, School of Life Sciences (ZBAF), University of Witten/Herdecke, D-58453 Witten, Germany., Knobloch J; Medical Clinic III for Pneumology, Allergology and Sleep Medicine, Bergmannsheil University Hospital, Ruhr-University Bochum, D-44789 Bochum, Germany., Park JJ; Department of Oto-Rhino-Laryngology-Head and Neck Surgery, St.-Josefs-Hospital Hagen, University of Witten/Herdecke, D-58097 Hagen, Germany. |
| Abstrakt: |
Epithelial cells may play an important role in the pathologic process of chronic rhinosinusitis with nasal polyps. Therefore, providing epithelial cells from a biobank could greatly contribute to further research. In the present work, the isolation of epithelial cells from long-term cryopreserved tissue is demonstrated. Polyp tissues were cryopreserved in a commercially available freezing medium with dimethyl sulfoxide and stored in liquid nitrogen. The outgrowth and proliferation of epithelial cells from cryopreserved tissue were evaluated and compared to that of fresh tissue. Flow cytometric analysis with anti-cytokeratin, anti-p63, and anti-Ki-67 was performed to identify epithelial cells and determine differentiation and proliferation. A functionality test was performed by determining type 2-relevant proteins, representatively thymic stromal lymphopoietin (TSLP) and periostin, using ELISA. Primary epithelial cells could be isolated from cryopreserved tissues. Cells from cryopreserved tissues showed comparable outgrowth and proliferation to that of fresh tissue. Isolated epithelial cells showed high cytokeratin, p63, and Ki-67 expression and secreted TSLP and periostin. In the present study, a method for long-term cryopreservation of polyp tissue was established, thereby enabling the isolation and cell culture of primary cell culture at a later time. Epithelial cell availability should be greatly improved by including this method in a biobank. |
