Human umbilical cord-derived mesenchymal stem cells (hucMSCs) can differentiate into multiple cell lineages, but few methods have been developed to generate kidney lineage cells. Due to their human origin, pluripotent nature and immunomodulatory properties, these stem cells are attractive candidates for clinical applications such as the repair or regeneration of damaged organs. This study evaluated the renal differentiation potential of hucMSCs, when exposed for 10 days to optimised concentrations of retinoic acid, activin-A and bone morphogenetic protein-7 (BMP-7) in various combinations, with and without the priming of the cells with a Wnt signalling pathway activator (CHIR99021). The hucMSCs were isolated and characterised according to surface marker expression (CD73, CD90, CD44, CD146 and CD8) and tri-lineage differentiation potential. The expression of key marker genes (OSR1, TBXT, HOXA13, SIX2, PAX2, KRT18 and ZO1) was examined by qRT-PCR. Specific marker protein expression (E-cadherin, cytokeratin-8 and cytokeratin-19) was analysed by immunocytochemistry. CHIR99021-primed cells treated with the retinoic acid, activin-A and BMP-7 cocktail showed epithelial cell-like differentiation — i.e. distinct phenotypic changes, as well as upregulated gene and protein expression, were observed that were consistent with an epithelial cell phenotype. Thus, our results showed that hucMSCs can efficiently differentiate into renal epithelial-like cells. This work may help in the development of focused therapeutic strategies, in which lineage-defined human stem cells can be used for renal regeneration.
WaikarSSLiuKDChertowGM. Diagnosis, epidemiology and outcomes of acute kidney injury. Clin J Am Soc Nephrol2008; 3: 844–861.
2.
KumarSLiuJMcMahonAP. Defining the renal repair transcriptome after acute kidney injury. Semin Nephrol2014; 34: 404–417.
3.
CheungCMPonnusamyAAndertonJG. Management of acute renal failure in the elderly patient: A clinician’s guide. Drugs Aging2008; 25: 455–476.
4.
WongCY. Current advances of stem cell-based therapy for kidney diseases. World J Stem Cells2021; 13: 914–933.
5.
DingDCChangYHShyuWC, et al.Human umbilical cord mesenchymal stem cells: A new era for stem cell therapy. Cell Transplant2015; 24: 339–347.
6.
MesserliMSJMarxCOppligerB, et al.Mesenchymal stem cells from Wharton’s jelly and amniotic fluid. Best Pract Res Clin Obstet Gynaecol2016; 31: 30–44.
7.
PengXXuHZhouY, et al.Human umbilical cord mesenchymal stem cells attenuate cisplatin-induced acute and chronic renal injury. Exp Biol Med2013; 238: 960–970.
8.
LiWYeBCaiXY, et al.Differentiation of human umbilical cord mesenchymal stem cells into prostate-like epithelial cells in vivo. PLoS One2014; 9: e102657.
9.
ShiQGaoJWJiangY, et al.Differentiation of human umbilical cord Wharton’s jelly-derived mesenchymal stem cells into endometrial cells. Stem Cell Res Ther2017; 8: 246.
10.
ChenHTangQLWuXY, et al.Differentiation of human umbilical cord mesenchymal stem cells into germ-like cells in mouse seminiferous tubules. Mol Med Rep2015; 12: 819–828.
11.
El-NagaAAEl-ChennawiFKamelSI, et al.The potentiality of human umbilical cord isolated mesenchymal stem/stromal cells for cardiomyocyte generation. Stem Cells Cloning2020; 13: 91–101.
12.
WangLDormerNHBonewaldLF, et al.Osteogenic differentiation of human umbilical cord mesenchymal stromal cells in polyglycolic acid scaffolds. Tissue Eng Part A2010; 16: 1937–1948.
13.
JinWXuYPYangAH, et al.In vitro induction and differentiation of umbilical cord mesenchymal stem cells into neuron-like cells by all-trans retinoic acid. Int J Ophthalmol2015; 8: 250–256.
14.
BrzoskaMGeigerHGauerS, et al.Epithelial differentiation of human adipose tissue-derived adult stem cells. Biochem Biophys Res Commun2005; 330: 142–150.
15.
KimDDresslerGR. Nephrogenic factors promote differentiation of mouse embryonic stem cells into renal epithelia. J Am Soc Nephrol2005; 16: 3527–3534.
16.
AraokaTSiMKuroseY, et al.Efficient and rapid induction of human iPSCs/ESCs into nephrogenic intermediate mesoderm using small molecule based differentiation methods. PLoS One2014; 9: e84881.
17.
NarayananKSchumacherKMTasnimF, et al.Human embryonic stem cells differentiate into functional renal proximal tubular-like cells. Kidney Int2013; 83: 593–603.
18.
YamaguchiSMorizaneRHommaK, et al.Generation of kidney tubular organoids from human pluripotent stem cells. Sci Rep2016; 6: 38353.
19.
WuYConnorsDBarberL, et al.Multiplexed assay panel of cytotoxicity in HK-2 cells for detection of renal proximal tubule injury potential of compounds. Toxicol In Vitro2009; 23: 1170–1178.
20.
TasnimFDengRHuM, et al.Achievements and challenges in bioartificial kidney development. Fibrogenesis Tissue Repair2010; 3: 14.
21.
HabibRHaneefKNaeemN, et al.Hypoxic stress and IL-7 gene overexpression enhance the fusion potential of rat bone marrow mesenchymal stem cells with bovine renal epithelial cells. Mol Cell Biochem2015; 403: 125–137.
PattersonLTPotterSS. Atlas of Hox gene expression in the developing kidney. Dev Dyn2004; 229: 771–779.
24.
LamAQFreedmanBSMorizaneR, et al.Rapid and efficient differentiation of human pluripotent stem cells into intermediate mesoderm that forms tubules expressing kidney proximal tubular markers. J Am Soc Nephrol2014; 25: 1211–1225.
25.
KuncewitchMYangWLCorboL, et al.WNT agonist decreases tissue damage and improves renal function after ischemia-reperfusion. Shock2015; 43: 268–275.
26.
ScottiMKherdjemilYRouxM, et al.A Hoxa13:Cre mouse strain for conditional gene manipulation in developing limb, hindgut, and urogenital system. Genesis2015; 53: 366–376.
27.
QinZZhouC. HOXA13 promotes gastric cancer progression partially via the FN1-mediated FAK/Src axis. Exp Hematol Oncol2022; 11: 7.
28.
RenXZhangJGongX, et al.Differentiation of murine embryonic stem cells toward renal lineages by conditioned medium from ureteric bud cells in vitro. Acta Biochim Biophys Sin (Shanghai)2010; 42: 464–471.
29.
BatchelderCALeeCCMatsellDG, et al.Renal ontogeny in the rhesus monkey (Macaca mulatta) and directed differentiation of human embryonic stem cells towards kidney precursors. Differentiation2009; 78: 45–56.
30.
ChenALiuYLuY, et al.Disparate roles of retinoid acid signaling molecules in kidney disease. Am J Physiol Renal Physiol2021; 320: F683–F692.
31.
ElbergGGuruswamySLoganCJ, et al.Plasticity of epithelial cells derived from human normal and ADPKD kidneys in primary cultures. Cell Tissue Res2008; 331: 495–508.
32.
ChenBXuXLinDD, et al.KRT18 modulates alternative splicing of genes involved in proliferation and apoptosis processes in both gastric cancer cells and clinical samples. Front Genet2021; 12: 635429.
33.
RahmanMSWruckWSpitzhornLS, et al.The FGF, TGF-β and WNT axis modulate self-renewal of human SIX2+ urine derived renal progenitor cells. Sci Rep2020; 10: 739.
34.
SanderVPrzepiorskiACrunkAE, et al.Protocol for large-scale production of kidney organoids from human pluripotent stem cells. STAR Protoc2020; 1: 100150.
35.
MugfordJWSipiläPMcMahonJA, et al.Osr1 expression demarcates a multi-potent population of intermediate mesoderm that undergoes progressive restriction to an Osr1-dependent nephron progenitor compartment within the mammalian kidney. Dev Biol2008; 324: 88–98.
36.
PengLHeQNLiXY, et al.Effect of homeobox A13 transfection on epithelial-mesenchymal transition and bone morphogenetic protein-7 expression in kidney tubular epithelial cells. Zhongguo Dang Dai Er Ke Za Zhi2015; 17: 1342–1347.
37.
BrunnerJRagupathySBorchardG. Target specific tight junction modulators. Adv Drug Deliv Rev2021; 171: 266–288.
38.
GriescheNHahnJBGeigerH, et al.During epithelial differentiation of human adipose-derived stromal/stem cells, expression of zonula occludens protein-1 is induced by a combination of retinoic acid, activin-A and bone morphogenetic protein-7. Cytotherapy2012; 14: 61–69.
39.
SelfMLagutinOVBowlingB, et al.Six2 is required for suppression of nephrogenesis and progenitor renewal in the developing kidney. EMBO J2006; 25: 5214–5228.
40.
JacobJTCoulombePAKwanR, et al.Types I and II keratin intermediate filaments. Cold Spring Harb Perspect Biol2018; 10: a018275.
