Non-invasive sources of cells with primary cilia from pediatric and adult patients

Cilia

Rachel H. Giles⁵  Michael A. Jewett¹  Marc R. Lilien⁴  Nine V. Knoers⁶  Mandy G. Keijzer-Veen⁴  Iris A. van Rooij²  Paulien A. Terhal⁶  Mieke M. van Haelst⁶  Kirsten Y. Renkema⁶  Hester Y. Kroes⁶  Ive Logister⁵  Heleen H. Arts³  Marijn F. Stokman⁶  Gisela G. Slaats⁵  Henry Ajzenberg⁵ 

[1] Department of Surgery (Urology), University of Toronto, 610 University Avenue, Toronto, M5G 2M9, Canada;Department of Health Evidence, Radboud University Medical Center, Geert Grooteplein zuid 10, Nijmegen, 6525GA, Netherlands;Department of Biochemistry, University of Western Ontario, Medical Sciences Building Rm. 342, Ontario, N6A 5C1, London, Canada;Department of Pediatric Nephrology, University Medical Center Utrecht, Lundlaan 6, Utrecht, 3584EA, Netherlands;Department of Nephrology and Hypertension, University Medical Center Utrecht, Heidelberglaan 100, Utrecht, 3584CX, Netherlands;Department of Medical Genetics, University Medical Center Utrecht, Lundlaan 6, Utrecht, 3584EA, Netherlands

关键词: Ciliopathy;    Cilia;    Protocol;    Cell culture;    Deciduous tooth;    Urine;    Pediatrics;   

Others  :  1210199
DOI  :  10.1186/s13630-015-0017-x

 received in 2015-01-27, accepted in 2015-05-05,  发布年份 2015

【 摘 要 】

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.

Findings

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.

【 授权许可】

   
2015 Ajzenberg et al.; licensee BioMed Central.

【 图 表 】

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【 参考文献 】

• [1]Waters AM, Beales PL: Ciliopathies: an expanding disease spectrum. Pediatr Nephrol 2011, 26:1039-56.
• [2]Hildebrandt F, Benzing T, Katsanis N: Ciliopathies. N Engl J Med 2011, 364:1533-43.
• [3]Zhou T, Benda C, Dunzinger S, Huang Y, Ho JC, Yang J, et al.: Generation of human induced pluripotent stem cells from urine samples. Nat Protoc 2012, 7:2080-9.
• [4]Giles RH, Ajzenberg H, Jackson PK: 3D spheroid model of mIMCD3 cells for studying ciliopathies and renal epithelial disorders. Nat Protoc 2014, 9:2725-31.
• [5]Hynes AM, Giles RH, Srivastava S, Eley L, Whitehead J, Danilenko M, et al.: Murine Joubert syndrome reveals Hedgehog signaling defects as a potential therapeutic target for nephronophthisis. Proc Natl Acad Sci U S A 2014, 111:9893-8.
• [6]Dorrenhaus A, Muller JI, Golka K, Jedrusik P, Schulze H, Follmann W: Cultures of exfoliated epithelial cells from different locations of the human urinary tract and the renal tubular system. Arch Toxicol 2000, 74:618-26.