Parallel generation of easily selectable multiple nephronal cell types from human pluripotent stem cells

Area of Science:

• Stem cell biology and regenerative medicine.
• Developmental biology and nephrogenesis.
• Biotechnology and bioprocess engineering.

Background:

• Human pluripotent stem cells (hPSCs) are a valuable source for generating various cell types.
• Generating defined kidney cells from hPSCs is crucial for regenerative medicine, disease modeling, and drug screening.
• Existing methods for hPSC-derived renal cells lack reproducibility, scalability, and efficiency.

Purpose of the Study:

• To establish a chemically defined, efficient, and scalable protocol for generating kidney cells from hPSCs.
• To produce specific renal cell populations, including metanephric mesenchyme (MM) and ureteric bud (UB)-like cells.
• To generate nephrogenic progenitors for downstream differentiation into various specialized kidney cell types.

Main Methods:

• Utilized a chemically defined protocol involving Activin A, BMP4, and Retinoic acid, followed by GDNF, to direct hPSCs towards the renal lineage.
• Characterized cell populations using immunofluorescence for SIX2, CITED1, HOXB7, and GRHL2.
• Performed transcriptome analysis to compare in vitro-derived cells with in vivo counterparts.

Main Results:

• Generated SIX2+/CITED1+ MM-like and HOXB7+/GRHL2+ UB-like cells within 6 days.
• Transcriptome analysis at day 8 confirmed resemblance to in vivo renal vesicle and ureteric epithelial cells.
• Developed tubular and glomerular renal cells by day 14, including mesangial, proximal tubular, distal tubular, collecting duct epithelial cells, and podocyte precursors.

Conclusions:

• The developed protocol rapidly and efficiently generates a pool of nephrogenic progenitors from hPSCs in 2D culture.
• This method provides a scalable basis for producing tailored renal cell types for therapeutic and research applications.
• The generated renal cells are suitable for drug testing and cell-based regenerative therapies.