J Appl Genetics (2011) 52:39–51 DOI 10.1007/s13353-010-0005-1 HUMAN GENETICS REVIEW In vitro culturing of ciliary respiratory cells—a model for studies of genetic diseases Zuzanna Bukowy & Ewa Ziętkiewicz & Michał Witt Received: 28 September 2010 / Accepted: 5 October 2010 / Published online: 2 December 2010 # The Author(s) 2010. This article is published with open access at Springerlink.com Abstract Primary ciliary dyskinesia (PCD) is a rare genetic disorder caused by the impaired functioning of ciliated cells. Its diagnosis is based on the analysis of the structure and functioning of cilia present in the respiratory epithelium (RE) of the patient. Abnormalities of cilia caused by hereditary mutations closely resemble and often overlap with defects induced by the environmental factors. As a result, proper diagnosis of PCD is difficult and may require repeated sampling of patients’ tissue, which is not always possible. The culturing of differentiated cells and tissues derived from the human RE seems to be the best way to diagnose PCD, to study genotype–phenotype relations of genes involved in ciliary dysfunction, as well as other aspects related to the functioning of the RE. In this review, different methods of culturing differentiated cells and tissues derived from the human RE, along with their potential and limitations, are summarized. Several consid- erations with respect to the factors influencing the process of in vitro differentiation (cell-to-cell interactions, medium composition, cell-support substrate) are also discussed. Keywords Respiratory epithelium . Cell culture . Cilia . Primary ciliary dyskinesia . Secondary ciliary dyskinesia Abbreviations AA Antibiotic-antimycotic solution BEGM Bronchial epithelial cell growth medium Z. Bukowy (*) : E. Ziętkiewicz : M. Witt Institute of Human Genetics, Polish Academy of Sciences, Poznań, Poland e-mail: zuza@man.poznan.pl Z. Bukowy : M. Witt International Institute of Molecular and Cell Biology, Warszawa, Poland CT DMEM EGF F12 HC IDA INS MT ODA PCD pen RA RE SCD strep TEM TR Cholera toxin Dulbecco’s modified Eagle’s medium Epithelial growth factor Ham’s F12 medium Hydrocortisone Inner dynein arms Insulin Microtubule Outer dynein arms Primary ciliary dyskinesia Penicillin Retinoic acid Respiratory epithelium Secondary ciliary dyskinesia Streptomycin Transmission electron microscopy Transferrin Primary ciliary dyskinesia Impaired functioning of cilia has been implicated in numerous human hereditary diseases, collectively referred to as ciliopathies. Their classification reflects the physio- logical role of different types of cilia involved, with the two main classes being motile cilia and sensory cilia (Badano et al. 2006). The human disorders due to the dysfunction of motile cilia are best exemplified by primary ciliary dyskinesia (PCD). PCD is a rare genetic disease (with a prevalence of 1:16,000 to 1:20,000 live births), characterized by recurrent respiratory airways infections, sinusitis, otitis media, bron- chiectasis, and male subfertility (Afzelius 1998; Schidlow 1994; Escudier et al. 2009). In about 50% of cases, PCD is