1. Directed differentiation of human induced pluripotent stem cells into mature kidney podocytes and establishment of a Glomerulus Chip
- Author
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Donald E. Ingber, Samira Musah, Nikolaos Dimitrakakis, Diogo M. Camacho, and George M. Church
- Subjects
0301 basic medicine ,Cellular differentiation ,Cytological Techniques ,Induced Pluripotent Stem Cells ,Kidney Glomerulus ,Microfluidics ,Kidney development ,Biology ,Regenerative medicine ,General Biochemistry, Genetics and Molecular Biology ,Article ,03 medical and health sciences ,0302 clinical medicine ,Directed differentiation ,Lab-On-A-Chip Devices ,medicine ,Humans ,Progenitor cell ,Induced pluripotent stem cell ,Glomerulus (olfaction) ,urogenital system ,Podocytes ,Cell Differentiation ,Cell biology ,030104 developmental biology ,medicine.anatomical_structure ,Cell culture ,030220 oncology & carcinogenesis - Abstract
Protocols have been established to direct the differentiation of human induced pluripotent stem (iPS) cells into nephron progenitor cells and organoids containing many types of kidney cells, but it has been difficult to direct the differentiation of iPS cells to form specific types of mature human kidney cells with high yield. Here, we describe a detailed protocol for the directed differentiation of human iPS cells into mature, post-mitotic kidney glomerular podocytes with high (>90%) efficiency within 26 days, and under chemically-defined conditions without genetic manipulations or subpopulation selection. We also describe how these iPS cell-derived podocytes may be induced to form within a microfluidic organ-on-a-chip (Organ Chip) culture device to build a human kidney Glomerulus Chip that mimics the structure and function of the kidney glomerular capillary wall in vitro within 35 days (starting with undifferentiated iPS cells). The podocyte differentiation protocol requires skills for culturing iPS cells, and the development of a Glomerulus Chip requires some experience with building and operating microfluidic cell culture systems. This method could be useful for applications in nephrotoxicity screening, therapeutic development, and regenerative medicine, as well as mechanistic study of kidney development and disease.
- Published
- 2018