DNA-stability and -ability of regeneration of human nasal mucosa in culture models

For culture models of primary cells of the human nasal mucosa, monocultures with epithelial cells (EC) are used as well as cocultures with EC and fibroblasts (FB). Furthermore, diverse methods of isolation exist. Although these techniques are established, no comparison and no evaluation of the relevance for experimental questions were conducted, yet.

EC and FB were isolated in 2 different methods (sequential growth of EC and FB respectively lysis and mechanical stripping). EC monocultures and compartment separated EC-FB cocultures were cultivated over three passages under air-liquid interface. EC and FB were separated immunobiologically from each other. DNA stability and regenerative capacity at DNA and chromosomal level as well as proliferation and cell differentiation were examined.

Both methods showed equivalent levels of DNA stability and regenerative capacity over all passages. After stripping the EC monoculture a passage depending contamination of FB was detected, but also a continuous proliferation. After sequential growth of the coculture a higher cell purity was observed within the compartment, but also a stagnation of growth in higher passages. Cilia were proved with electron microscopy in both methods. The isolation technique of sequential growth generates purer cell populations of FB and EC. The following coculture under air-liquid interface enables conditions close to in vivo situation, because of the cell cell communication. That's why this procedure is more accurate and is the method of choice in experimental rhinology, although the duration of cultivation is limited and the DNA stability in both methods is equal.

Publication History

Publication Date:
23 April 2019 (online)