Bioengineering Renal Epithelial-Like Cells from Mesenchymal Stem Cells by Combinations of Growth Factors and Small Molecules

This research aimed to differentiate mouse adipose-derived mesenchymal stem cells (mAd-MSCs) into renal epithelial-like cells.

Three treatments (T1, T2, and T3), with combinations of growth factors and small molecules, were used in stem cell signature mAd-MSCs in a defined serum-free culture medium. The growth factors and chemicals included were HGF, IGF, PDGF, FGF2, Bmp7, and RepSox, CHIR99021, respectively. The reprogrammed cells and their respective controls were evaluated by transcriptional and translational analyses. Albumin binding and uptake co-localization assays and co-culturing of differentiated cells with murine adult renal cortical cells were also observed.

The inductive signals via three treatments using growth factors and small molecules produced morphological dynamics from spindle shape to squared shape epithelial-like cells. The molecular analysis revealed elevated expressions of Pdpn, Cd2ap/Mets1, Agt. Jag1, Erk2/Mapk, and P-cadherin/Cdh3 with low gene expression of Tgf-β1. Nphs2/Podocin and Ksp/Cdh16 were expressed in protein analyses of differentiated cells, while CD44 was declined. Functionally, the differentiated cells exhibit albumin endocytosis via megalin receptor expression. The fusion of reprogrammed cells with murine cortical renal epithelial cells presented further connections, integration stability, and growth of cells for up to 10 days.

Our novel strategy of reprogramming displayed efficient commitments of MSCs toward renal epithelial-like cells with not only an orchestration of gene and protein expressions but also functional commitment. The study supplemented information for the ex-vivo reprogramming of MSCs into renal epithelialization, which led to the development of a novel regenerative approach.

Kidney diseases are increasing worldwide and have become the leading cause of death. The regeneration capabilities of kidneys are up to an extent, and insufficiency leads to hemodialysis in kidney patients, especially for chronic kidney diseases, as a therapeutic option. Currently, the gold standard is kidney transplantation, for which a donor kidney must be required. The donor’s kidney is lacking, and currently, no functional artificial organ is available. Several stem cell approaches are being investigated for a better treatment option through simple or modified cells to restore kidney homeostasis. This study is to investigate the role of growth factors and small molecules in combination, for renal epithelial-like cell generation from an easy source of stem cells, i.e., adipose-derived mesenchymal stem cells. The reprogramming approach proved successful, and it not only induced differentiation but also exhibited gene and protein expression of functional attributes. The cells displayed morphological variability from spindle to a prominent typical cobblestone shape. They have the capability of albumin uptake via megalin receptors and have physiologic integration capability with renal epithelial cells, which was tested for up to 10 days.