Induced Pluripotent Stem Cell (iPSc)-Derived and Primary Podocyte-Like Cells Are Capable of Macromolecular Sieving After Culture in Scalable Scaffolds
- Bioengineering, Augmented Intelligence, Digital Health, and Data Science
November 04, 2023 | Location: Room 108, Pennsylvania Convention Center
Abstract Time: 04:39 PM - 04:48 PM
- 400 Bioengineering
- Gallegos, Thomas F., Iviva Medical, Woburn, Massachusetts, United States
- Przepiorski, Aneta J., Iviva Medical, Woburn, Massachusetts, United States
- Sarikhani, Mohsen, Iviva Medical, Woburn, Massachusetts, United States
With the growing demand for donor kidneys and insufficient donor organs, there is an urgent need for alternatives to dialysis and kidney transplantation. Our team has developed a biocompatible engineered glomerular tissue construct comprising hollow vascular and epithelial networks separated by a porous biomimetic membrane. The scaffolds are seeded with endothelial cells and podocytes that when perfused under vascular pressure, recapitulate glomerular function to produce a primary filtrate. The glomerular units are seeded with primary cells isolated from non-transplantable human donor kidneys of by isolation of cells from human induced pluripotent stem cells (iPSCs), that have been differentiated into endothelial cells and podocyte-like cells. We evaluated filtration dynamics and cell characteristics.
Human primary podocytes and endothelial cells are isolated by enzymatic digestion and glomerular isolation by sieving. Podocytes and endothelial cells were isolated by subsequent glomerular digestion and FACS sorting. Isolates were perfusion seeded. Human iPSCs were differentiated into endothelium using an established protocol. Human kidney organoids were generated using published methods with modifications. MAFB positive podocyte clusters were isolated using enzyme digestion and size filtration and expanded to form a monolayer. Cell seeded scaffolds were cultured using custom bioreactors and perfusion control units. Physiology was assayed by vascular perfusion under defined backpressure to generate primary filtrate through the epithelial channel.
Acellular control scaffolds are highly permissive to transmembrane fluid flux and produce filtrates which contain micro- and macromolecules. Histological analysis of fixed primary and iPSC scaffolds shows cellular coverage within the endothelial and podocyte channels, indicating good cellular attachment and survival under perfusion for a minimum of 7 days. Pressurized perfusion of cellularized scaffolds with a blood-like buffer containing physiologic albumin and hematocrit through the vascular channel, produced RBC-free filtrate which resists macromolecule loss from the vasculature.
Our results demonstrate that both primary and iPSc derived endothelial and podocyte-like cells are a viable option for physiologically relevant engineered renal tissues.
- Commercial Support – Iviva Medical