Investigation of Cellular Function and DNA Integrity during 2D in vitro Culture of Human Salivary Gland Epithelial Cells.


Journal

Cells, tissues, organs
ISSN: 1422-6421
Titre abrégé: Cells Tissues Organs
Pays: Switzerland
ID NLM: 100883360

Informations de publication

Date de publication:
Historique:
received: 27 09 2019
accepted: 15 12 2019
pubmed: 6 2 2020
medline: 6 8 2020
entrez: 6 2 2020
Statut: ppublish

Résumé

In vitro culture of human salivary gland epithelial cells (SGEC) is still a challenge. A high quantity and quality of cells are needed for the cultivation of 3D matrices. Furthermore, it is known that DNA damage is supposed to be an important factor involved in carcinogenesis. This study investigates cellular function and DNA integrity of human SGEC during 3 passage steps in 2 groups (group 1: n = 10; group 2: n = 9). Cellular function was analyzed by immunofluorescence, transmission electron microscopy (TEM), and quantitative real-time polymerase chain reaction (qPCR). DNA integrity was tested via the comet assay. Immunohistochemistry and qPCR results showed stable α-amylase and pan-cytokeratin levels; TEM revealed functional cells; and no significant DNA damage could be detected in the comet assay during 3 culture steps. The study shows that not only at cellular but also at DNA level human SGEC can be safely quantified over 3 passages for preclinical tissue engineering without loss of differentiation and function.

Identifiants

pubmed: 32023622
pii: 000505433
doi: 10.1159/000505433
doi:

Substances chimiques

Keratins 68238-35-7
alpha-Amylases EC 3.2.1.1

Types de publication

Journal Article

Langues

eng

Sous-ensembles de citation

IM

Pagination

66-75

Informations de copyright

© 2020 S. Karger AG, Basel.

Auteurs

Marc Burghartz (M)

Department of Otorhinolaryngology, Head and Neck Surgery, Klinikum Stuttgart, Stuttgart, Germany.

Johannes Taeger (J)

Department of Otorhinolaryngology, Plastic, Aesthetic, and Reconstructive Head and Neck Surgery, University Hospital Würzburg, Würzburg, Germany, taeger_j@ukw.de.

Marco Metzger (M)

Department of Tissue Engineering and Regenerative Medicine (TERM), University Hospital Würzburg, Würzburg, Germany.

Agmal Scherzad (A)

Department of Otorhinolaryngology, Plastic, Aesthetic, and Reconstructive Head and Neck Surgery, University Hospital Würzburg, Würzburg, Germany.

Thomas Gehrke (T)

Department of Otorhinolaryngology, Plastic, Aesthetic, and Reconstructive Head and Neck Surgery, University Hospital Würzburg, Würzburg, Germany.

Pascal Ickrath (P)

Department of Otorhinolaryngology, Plastic, Aesthetic, and Reconstructive Head and Neck Surgery, University Hospital Würzburg, Würzburg, Germany.

Evelyn Kolb (E)

Department of Otorhinolaryngology, Plastic, Aesthetic, and Reconstructive Head and Neck Surgery, University Hospital Würzburg, Würzburg, Germany.

Norbert Kleinsasser (N)

Department of Otorhinolaryngology, Head and Neck Surgery, Kepler University, Linz, Austria.

Rudolf Hagen (R)

Department of Otorhinolaryngology, Plastic, Aesthetic, and Reconstructive Head and Neck Surgery, University Hospital Würzburg, Würzburg, Germany.

Stephan Hackenberg (S)

Department of Otorhinolaryngology, Plastic, Aesthetic, and Reconstructive Head and Neck Surgery, University Hospital Würzburg, Würzburg, Germany.

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Classifications MeSH