Primary ciliary dyskinesia (PCD) is a congenital disease leading to an impairment of the mucociliary clearance.Culturing primary human epithelial cells (phNEC) at air-liquid-interface (ALI) is the standard model for diagnostics and functional characterization.Challenges are a low yield and short lifespan of phNEC.A conditionally reprogrammed cell (CRC) system could overcome this, by enhancing cell growth and lifespan, preserving the cell-of-origin function as shown for primary bronchial epithelial cells(1).Therefore, we aimed to establish and optimize the CRC technology for phNEC from people with PCD (pwPCD). Nasal brushings were taken from 29pwPCD with genetically confirmed diagnosis. phNEC were cultured with immortalized ^3T3fibroblasts in the presence of Y-27632.After expansion, cells were partially cryopreserved (biobanked), further passaged and cultured at ALI.Ciliary Beat Frequency (CBF) was assessed prior to culturing and at ALI.Morphology and cell type composition were determined by immunofluorescence staining (IF). Utilizing the CRC method for patient-derived phNEC had an excellent success rate (>90%) and high cell count yield (median:10x10⁶ cells). CBF showed comparable results in freshly obtained vs cells at ALI. Disturbed mucociliary clearance was retained and IF staining revealed pseudostratified, ciliated epithelial layers in differentiated ALI cultures.Utilizing the CRC technology for phNEC from pwPCD stands out by its non-invasiveness, excellent success rate and high cell yield, which allows successful biobanking. This method can be used for functional characterization as well as high throughput screening for therapeutic targets as a step towards personalized medicine for pwPCD