Neuhaus, V.V.NeuhausSchaudien, D.D.SchaudienGolovina, T.T.GolovinaTemann, U.-A.U.-A.TemannThompson, C.C.ThompsonLippmann, T.T.LippmannBersch, C.C.BerschPfennig, O.O.PfennigJonigk, D.D.JonigkBraubach, P.P.BraubachFieguth, H.-G.H.-G.FieguthWarnecke, G.G.WarneckeYusibov, V.V.YusibovSewald, K.K.SewaldBraun, A.A.Braun2022-03-052022-03-052017https://publica.fraunhofer.de/handle/publica/24904210.1186/s12995-017-0158-5Background: Investigation of basic chronic inflammatory mechanisms and development of new therapeutics targeting the respiratory tract requires appropriate testing systems, including those to monitor long- persistence. Human precision-cut lung slices (PCLS) have been demonstrated to mimic the human respiratory tract and have potential of an alternative, ex-vivo system to replace or augment in-vitro testing and animal models. So far, most research on PCLS has been conducted for short cultivation periods (â¤72 h), while analyses of slowly metabolized therapeutics require long-term survival of PCLS in culture. In the present study, we evaluated viability, physiology and structural integrity of PCLS cultured for up to 15 days. Methods: PCLS were cultured for 15 days and various parameters were assessed at different time points. Results: Structural integrity and viability of cultured PCLS remained constant for 15 days. Moreover, bronchoconstriction was inducible over the wh ole period of cultivation, though with decreased sensitivity (EC501d = 4 à 10-8 M vs. EC5015d = 4 à 10-6 M) and reduced maximum of initial airway area (1d = 0.5% vs. 15d = 18.7%). In contrast, even though still clearly inducible compared to medium control, LPS-induced TNF-α secretion decreased significantly from day 1 to day 15 of culture. Conclusions: Overall, though long-term cultivation of PCLS need further investigation for cytokine secretion, possibly on a cellular level, PCLS are feasible for bronchoconstriction studies and toxicity assays.en610Assessment of long-term cultivated human precision-cut lung slices as an ex vivo system for evaluation of chronic cytotoxicity and functionalityjournal article