In vitro culturing of porcine tracheal mucosa as an ideal model for investigating the influence of drugs on human respiratory mucosa

Abstract

It has been previously shown that fresh mucosa from diVerent mammals could serve as raw material for in vitro culturing with the diVerentiation of cilia, which are the most important morphological structures for the function of the mucociliary system. Increasing legal restrictions on the removal of human tissue and changing surgical techniques have led to a lack of fresh human mucosa for culturing. Most of the animals that have been used as donors up to now are genetically not very close to human beings and must all be sacriWced for such studies. We, therefore, established a modiWed system of culturing mucosa cells from the trachea of pigs, which is available as a regular by-product after slaughtering. With respect to the possibility of developing "beating" cilia, it could be shown that the speed of cell proliferation until adhesion to the coated culture dishes, the formation of conjunctions of cell clusters and the proliferation of cilia were comparable for porcine and human mucosa. Moreover, it could be demonstrated that the porcine cilia beat frequency of 7.57 § 1.39 Hz was comparable to the human mucosa cells beat frequency of 7.3 § 1.4 Hz and that this beat frequency was absolutely constant over the investigation time of 360 min. In order to prove whether the reaction to diVerent drugs is comparable between the porcine and human cilia, we initially tested benzalkonium chloride, which is known to be toxic for human cells, followed by naphazoline, which we found in previous studies on human mucosa to be non-toxic. The results clearly showed that the functional and morphological reactions of the porcine ciliated cells to these substances were similar to the reaction we found in the in vitro cultured human mucosa.

Ciliated Air-Liquid Cultures as an Aid to Diagnostic Testing of Primary Ciliary Dyskinesia

“…13 Also, unlike our current study, the precise ciliary beat pattern was not evaluated. 13 Both the air-liquid interface [23][24][25] and submerged cell culture techniques have proved to be useful in research relating to respiratory disease. Of note, the submerged culture technique has recently been used to demonstrate that normal ciliary movement can be restored by lentiviral gene therapy.…”

Rock Inhibitor Y-27632 Enables Feeder-Free, Unlimited Expansion of Sus scrofa domesticus Swine Airway Stem Cells to Facilitate Respiratory Research

“…Despite the need for a readily available cell source to bridge the gap between laboratory research and application, little work has been carried out to develop AEC culture with large, nonprimate mammals. Previous work over the last four decades, though limited, has focussed on dogs [25–27], sheep [28–33], horses [34], cows [35–39], and pigs [9, 27, 40–44] with little to no cell expansion being a common feature, as nonruminant omnivore pigs may prove to have the most similar biology to humans with comparable respiratory characteristics [45]. Here, we have successfully isolated and cultured mucociliary differentiation-competent pAECs that can reproducibly yield high, potentially unlimited, cells.…”

Research on capacity addition using market model with transmission congestion under competitive environment

“…ATP is a well‐known cilio‐stimulating drug,10 while BAC is 1 of the most commonly used ciliotoxic drugs. The concentration of 0.005% BAC caused a significant decrease of CBF both in cultured human nasal and porcine tracheal epithelial cells 11. In the present study, we compared the ciliary reactivity of mouse nasal septal and turbinal epithelial cells in response to different concentrations of ATP or BAC.…”