3D polyelectrolyte scaffolds to mimic exocrine glands: a step towards a prostate-on-chip platform

Open access


We report our approach to creating a microfluidic chip (namely UroLOC) that mimics the acinar/tubular structure and the luminal microenvironment of exocrine glands. The chip utilises a nanostructured membrane that is designed to provide a 3-dimensional supporting scaffold for the growth of exocrine acinus epithelial cells. The nanostructured membrane was produced using layer-by-layer assembly of polyelectrolytes, and formed into 3-dimensional hemispherical cavities and “finger-like” structures in order to mimic the natural architecture of acini found in exocrine glands. We utilised normal (PNT2) and cancerous (PC3, LNCaP) prostate epithelial cells to demonstrate the proof-of-concept of using MALDI (Matrix Assisted Laser Desorption Ionisation) profiling of secretions collected after 48 hours of cell growth, with no concentration or purification steps and without any a priori on the knowledge of targeted proteins. This MALDI profiling analysis of the crude supernatants from 3 different cell lines (PNT2, PC3 and LNCaP) demonstrated the capacity of the MALDI profiling approach to discriminate between the different secretome signatures. The UroLOC concept and secretome profiling that we describe opens new opportunities in terms of liquid-biopsy based diagnosis, particularly for the early stages of carcinogenesis.

1. Vidi P-A, Bissell MJ, Lelievre SA. Three-Dimensional Culture of Human Breast Epithelial Cells: The How and the Why. Methods Mol Biol. 2013;945:193-219.

2. Mosaad EO, Chambers KF, Futrega K, Clements JA, Doran MR. The Microwell-mesh: A high-throughput 3D prostate cancer spheroid and drug-testing platform. Sci Rep. 10 janv 2018;8(1):253.

3. Picollet-D’hahan N, Dolega ME, Freida D, Martin DK, Gidrol X. Deciphering Cell Intrinsic Properties: A Key Issue for Robust Organoid Production. Trends Biotechnol. 2017;35(11):1035-48.

4. Picollet-D’hahan N, Dolega ME, Liguori L, Marquette C, Le Gac S, Gidrol X, et al. A 3D Toolbox to Enhance Physiological Relevance of Human Tissue Models. Trends Biotechnol. 2016;34(9):757-69.

5. Kim HJ, Ingber DE. Gut-on-a-Chip microenvironment induces human intestinal cells to undergo villus differentiation. Integrative Biology. 2013;5(9):1130.

6. Kim HJ, Li H, Collins JJ, Ingber DE. Contributions of microbiome and mechanical deformation to intestinal bacterial overgrowth and inflammation in a human gut-on-a-chip. Proceedings of the National Academy of Sciences. 5 janv 2016;113(1):E7-15.

7. Costello CM, Hongpeng J, Shaffiey S, Yu J, Jain NK, Hackam D, et al. Synthetic small intestinal scaffolds for improved studies of intestinal differentiation: Synthetic Small Intestinal Scaffolds for Improved. Biotechnology and Bioengineering. juin 2014;111(6):1222-32.

8. Wang Y, Gunasekara DB, Reed MI, DiSalvo M, Bultman SJ, Sims CE, et al. A microengineered collagen scaffold for generating a polarized crypt-villus architecture of human small intestinal epithelium. Biomaterials. 1 juin 2017;128:44-55.

9. Bein A, Shin W, Jalili-Firoozinezhad S, Park MH, Sontheimer-Phelps A, Tovaglieri A, et al. Microfluidic Organ-on-a-Chip Models of Human Intestine. Cell Mol Gastroenterol Hepatol. 24 avr 2018;5(4):659-68.

10. Battle AR, Valenzuela SM, Mechler AI, Nichols RJ, Praporski S, Maio IL di, et al. Novel engineered ion channel provides controllable ion permeability for polyelectrolyte microcapsules coated with a lipid membrane. Advanced Functional Materials. 2009;19:201-8.

11. Picollet-D’hahan N, Gerbaud S, Kermarrec F, Alcaraz J-P, Obeid P, Bhajun R, et al. The modulation of attachment, growth and morphology of cancerous prostate cells by polyelectrolyte nanofilms. Biomaterials. dec 2013;34(38):10099-108.

12. Debnath J, Brugge JS. Modelling glandular epithelial cancers in three-dimensional cultures. Nat Rev Cancer. sept 2005;5(9):675-88.

13. Dolega ME, Wagh J, Gerbaud S, Kermarrec F, Alcaraz J-P, Martin DK, et al. Facile Bench-Top Fabrication of Enclosed Circular Microchannels Provides 3D Confined Structure for Growth of Prostate Epithelial Cells. Acott TS, editeur. PLoS ONE. 19 juin 2014;9(6):e99416.

14. Sung JH, Yu J, Luo D, Shuler ML, March JC. Microscale 3-D hydrogel scaffold for biomimetic gastrointestinal (GI) tract model. Lab Chip. 7 fevr 2011;11(3):389-92.

15. Kim HJ, Ingber DE. Gut-on-a-Chip microenvironment induces human intestinal cells to undergo villus differentiation. Integr Biol (Camb). sept 2013;5(9):1130-40.

16. Lang SH, Sharrard RM, Stark M, Villette JM, Maitland NJ. Prostate epithelial cell lines form spheroids with evidence of glandular differentiation in three-dimensional Matrigel cultures. Br J Cancer. 17 aout 2001;85(4):590-9.

17. Francis GL. Albumin and mammalian cell culture: implications for biotechnology applications. Cytotechnology. janv 2010;62(1):1-16.

18. EVECEN M, PABUCCUO S. The Effects of Various BSA Levels in Different Media on Development in In Vitro Culture of Mouse Embryos.. K. :6.

19. Peter JF, Otto AM, Wolf B. Enrichment and Detection of Molecules Secreted by Tumor Cells Using Magnetic Reversed-Phase Particles and LC-MALDI-TOF-MS. 2007;18(5):11.

20. Godugu C, Patel AR, Desai U, Andey T, Sams A, Singh M. AlgiMatrixTM based 3D cell culture system as an in-vitro tumor model for anticancer studies. PLoS ONE. 2013;8(1):e53708.

21. Fontenete S, Silva J, Teixeira AL, Ribeiro R, Bastos E, Pina F, et al. Controversies in using urine samples for Prostate Cancer detection: PSA and PCA3 expression analysis. Int Braz J Urol. dec 2011;37(6):719-26.

Journal Information


All Time Past Year Past 30 Days
Abstract Views 0 0 0
Full Text Views 155 155 59
PDF Downloads 123 123 29