title: Non-invasive sources of cells with primary cilia from pediatric and adult patients.
abstract: BACKGROUND
  Ciliopathies give rise to a multitude of organ-specific pathologies; obtaining relevant primary patient material is useful for both diagnostics and research. However, acquisition of primary ciliated cells from patients, particularly pediatric patients, presents multiple difficulties. Biopsies and blood samples are invasive, and patients (and their parents) may be reluctant to travel to medical centers, especially for research purposes. We sought to develop non-invasive methods of obtaining viable and ciliated primary cells from ciliopathy patients which could be obtained in the home environment.
  
  
  RESULTS
  We introduce two methods for the non-invasive acquisition of primary ciliated cells. In one approach, we collected spontaneously shed deciduous (milk) teeth from children. Fibroblast-like cells were observed after approximately 2 weeks of culture of fragmented teeth. Secondly, urine samples were collected from children or adults. Cellular content was isolated and after approximately 1 week, renal epithelial cells were observed. Both urine and tooth-derived cells ciliate and express ciliary proteins visible with immunofluorescence. Urine-derived renal epithelial cells (URECs) are amenable to 3D culturing, siRNA knockdown, and ex vivo drug testing.
  
  
  CONCLUSIONS
  As evidence continues to accumulate showing that the primary cilium has a central role in development and disease, the need for readily available and ciliated patient cells will increase. Here, we introduce two methods for the non-invasive acquisition of cells with primary cilia. We believe that these cells can be used for further ex vivo study of ciliopathies and in the future, for personalized medicine.
authors.affiliation: Department of Nephrology and Hypertension, University Medical Center Utrecht, Heidelberglaan 100, 3584CX Utrecht, Netherlands.
authors.forename: Henry
authors.initials: H
authors.lastname: Ajzenberg
authors.affiliation: Department of Nephrology and Hypertension, University Medical Center Utrecht, Heidelberglaan 100, 3584CX Utrecht, Netherlands.
authors.forename: Gisela G
authors.initials: GG
authors.lastname: Slaats
authors.affiliation: Department of Medical Genetics, University Medical Center Utrecht, Lundlaan 6, 3584EA Utrecht, Netherlands.
authors.forename: Marijn F
authors.initials: MF
authors.lastname: Stokman
authors.affiliation: Department of Human Genetics, Radboud University Medical Center, Geert Grooteplein zuid 10, 6525GA Nijmegen, Netherlands.|Department of Biochemistry, University of Western Ontario, Medical Sciences Building Rm. 342, N6A 5C1 London Ontario, Canada.
authors.forename: Heleen H
authors.initials: HH
authors.lastname: Arts
authors.affiliation: Department of Nephrology and Hypertension, University Medical Center Utrecht, Heidelberglaan 100, 3584CX Utrecht, Netherlands.
authors.forename: Ive
authors.initials: I
authors.lastname: Logister
authors.affiliation: Department of Medical Genetics, University Medical Center Utrecht, Lundlaan 6, 3584EA Utrecht, Netherlands.
authors.forename: Hester Y
authors.initials: HY
authors.lastname: Kroes
authors.affiliation: Department of Medical Genetics, University Medical Center Utrecht, Lundlaan 6, 3584EA Utrecht, Netherlands.
authors.forename: Kirsten Y
authors.initials: KY
authors.lastname: Renkema
authors.affiliation: Department of Medical Genetics, University Medical Center Utrecht, Lundlaan 6, 3584EA Utrecht, Netherlands.
authors.forename: Mieke M
authors.initials: MM
authors.lastname: van Haelst
authors.affiliation: Department of Medical Genetics, University Medical Center Utrecht, Lundlaan 6, 3584EA Utrecht, Netherlands.
authors.forename: Paulien A
authors.initials: PA
authors.lastname: Terhal
authors.affiliation: Department of Health Evidence, Radboud University Medical Center, Geert Grooteplein zuid 10, 6525GA Nijmegen, Netherlands.
authors.forename: Iris A
authors.initials: IA
authors.lastname: van Rooij
authors.affiliation: Department of Pediatric Nephrology, University Medical Center Utrecht, Lundlaan 6, 3584EA Utrecht, Netherlands.
authors.forename: Mandy G
authors.initials: MG
authors.lastname: Keijzer-Veen
authors.affiliation: Department of Medical Genetics, University Medical Center Utrecht, Lundlaan 6, 3584EA Utrecht, Netherlands.
authors.forename: Nine V
authors.initials: NV
authors.lastname: Knoers
authors.affiliation: Department of Pediatric Nephrology, University Medical Center Utrecht, Lundlaan 6, 3584EA Utrecht, Netherlands.
authors.forename: Marc R
authors.initials: MR
authors.lastname: Lilien
authors.affiliation: Department of Surgery (Urology), University of Toronto, 610 University Avenue, M5G 2M9 Toronto, Canada.
authors.forename: Michael A
authors.initials: MA
authors.lastname: Jewett
authors.affiliation: Department of Nephrology and Hypertension, University Medical Center Utrecht, Heidelberglaan 100, 3584CX Utrecht, Netherlands.
authors.forename: Rachel H
authors.initials: RH
authors.lastname: Giles
keywords: Cell culture; Cilia; Ciliopathy; Deciduous tooth; Pediatrics; Protocol; Urine