Curvature facilitates podocyte culture in a biomimetic platform
| Autor: | Samad Ahadian, Erding Hu, Christian H. James, Nathaniel P. Smith, Irene Gu, Robert N. Willette, Claire Velikonja, Boyang Zhang, Milica Radisic, Anastasia Korolj, Carol Laschinger |
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| Rok vydání: | 2018 |
| Předmět: |
0301 basic medicine
Cell Cell Culture Techniques Biomedical Engineering Bioengineering 02 engineering and technology Biochemistry Podocyte Nephrin Mice 03 medical and health sciences Biomimetics In vivo Gene expression medicine Animals Barrier function biology Podocytes Chemistry Membrane Proteins General Chemistry 021001 nanoscience & nanotechnology Transport protein Cell biology Protein Transport 030104 developmental biology medicine.anatomical_structure Cell culture biology.protein 0210 nano-technology |
| Zdroj: | Lab on a Chip. 18:3112-3128 |
| ISSN: | 1473-0189 1473-0197 |
| DOI: | 10.1039/c8lc00495a |
| Popis: | Most kidney diseases begin with abnormalities in glomerular podocytes, motivating the need for podocyte models to study pathophysiological mechanisms and new treatment options. However, podocytes cultured in vitro face a limited ability to maintain appreciable extents of differentiation hallmarks, raising concerns over the relevance of study results. Many key properties such as nephrin expression and morphology reach plateaus that are far from the in vivo levels. Here, we demonstrate that a biomimetic topography, consisting of microhemispheres arrayed over the cell culture substrate, promotes podocyte differentiation in vitro. We define new methods for fabricating microscale curvature on various substrates, including a thin porous membrane. By growing podocytes on our topographic substrates, we found that these biophysical cues augmented nephrin gene expression, supported full-size nephrin protein expression, encouraged structural arrangement of F-actin and nephrin within the cell, and promoted process formation and even interdigitation compared to the flat substrates. Furthermore, the topography facilitated nephrin localization on curved structures while nuclei lay in the valleys between them. The improved differentiation was also evidenced by tracking barrier function to albumin over time using our custom topomembranes. Overall, our work presents accessible methods for incorporating microcurvature on various common substrates, and demonstrates the importance of biophysical stimulation in supporting higher-fidelity podocyte cultivation in vitro. |
| Databáze: | OpenAIRE |
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