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Transplantation of induced pluripotent stem cell-derived renal stem cells improved acute kidney injury.

Li Q, Tian SF, Guo Y, Niu X, Hu B, Guo SC, Wang NS, Wang Y - Cell Biosci (2015)

Bottom Line: However, the therapeutic effect of iPSC-derived RPCs for AKI has yet to be determined.We then established the rat ischemia-reperfusion injury (IR) model and transplanted the iPSC-derived RPCs into the injured rats in combination with the hydrogel.Our results revealed that iPSC-derived RPCs can protect AKI rat from renal function impairment and severe tubular injury by up-regulating the renal tubules formation, promoting cell proliferation, reducing apoptosis, and regulating the microenvironment in the injured kidney.

View Article: PubMed Central - PubMed

Affiliation: Institute of Microsurgery on Extremities, Shanghai Jiao Tong University Affiliated Sixth People's Hospital, Shanghai, 200233 China.

ABSTRACT

Background: Acute kidney injury (AKI) is a severe disease with high morbidity and mortality. Methods that promote repair of the injured kidney have been extensively investigated. Cell-based therapy with mesenchymal stem cells or renal progenitor cells (RPCs) resident in the kidney has appeared to be an effective strategy for the treatment of AKI. Embryonic stem cells or induced pluripotent stem cells (iPSCs) are also utilized for AKI recovery. However, the therapeutic effect of iPSC-derived RPCs for AKI has yet to be determined.

Methods: In this study, we induced iPSCs differentiation into RPCs using a nephrogenic cocktail of factors combined with the renal epithelial cell growth medium. We then established the rat ischemia-reperfusion injury (IR) model and transplanted the iPSC-derived RPCs into the injured rats in combination with the hydrogel. Next, we examined the renal function-related markers and renal histology to assess the therapeutic effect of the injected cells. Moreover, we investigated the mechanism by which iPSC-derived RPCs affect AKI caused by IR.

Results: We showed that the differentiation efficiency of iPSCs to RPCs increased when cultured with renal epithelial cell growth medium after stimulation with a nephrogenic cocktail of factors. The transplantation of iPSC-derived RPCs decreased the levels of biomarkers indicative of renal injury and attenuated the necrosis and apoptosis of renal tissues, but resulted in the up-regulation of renal tubules formation, cell proliferation, and the expression of pro-renal factors.

Conclusion: Our results revealed that iPSC-derived RPCs can protect AKI rat from renal function impairment and severe tubular injury by up-regulating the renal tubules formation, promoting cell proliferation, reducing apoptosis, and regulating the microenvironment in the injured kidney.

No MeSH data available.


Related in: MedlinePlus

Efficient differentiation of renal progenitor cells from mouse induced pluripotent stem cells (iPSCs). The renal differentiation of mouse iPSCs was initiated by the formation of embryoid bodies (EBs). After 2 days, retinoic acid, activin A, and BMP7 were added to the differentiation medium, and the cells were exposed to this supplemented media for 5 days (RAB group). After incubation with growth factors for 5 days, the EBs in the RAB + REGM group were cultured in renal epithelial cell growth medium for another 5 days. a Real-time PCR analysis of the expression of mesoderm (Bry) and renal lineage genes (Pax2, Osr1, WT1, Six2, CD24, Sal1, AQP1, E-cad and PDGFR). b Flow cytometry analysis of the expression of mesoderm renal progenitor-related genes. Black line, cells stained with isotype control; Red line, cells stained with indicated antibodies. *P < 0.05
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Fig1: Efficient differentiation of renal progenitor cells from mouse induced pluripotent stem cells (iPSCs). The renal differentiation of mouse iPSCs was initiated by the formation of embryoid bodies (EBs). After 2 days, retinoic acid, activin A, and BMP7 were added to the differentiation medium, and the cells were exposed to this supplemented media for 5 days (RAB group). After incubation with growth factors for 5 days, the EBs in the RAB + REGM group were cultured in renal epithelial cell growth medium for another 5 days. a Real-time PCR analysis of the expression of mesoderm (Bry) and renal lineage genes (Pax2, Osr1, WT1, Six2, CD24, Sal1, AQP1, E-cad and PDGFR). b Flow cytometry analysis of the expression of mesoderm renal progenitor-related genes. Black line, cells stained with isotype control; Red line, cells stained with indicated antibodies. *P < 0.05

Mentions: To investigate whether iPSC-derived RPCs can improve AKI, we first used a nephrogenic cocktail of factors (including retinoic acid, activin A, and BMP7) to induce iPSCs differentiation into RPCs (Additional file 1: Figure S1). The iPS cell line used in this study is tagged with a ubiquitously expressed gfp gene. The real-time PCR analysis showed that the expression of the RPC markers PAX2, WT1, Six2 and CD24 increased after RAB treatment (Fig. 1a), indicating that RPCs were successfully induced. As shown by flow cytometry analysis, the PAX2, WT1 and CD24 positive cells were 33.16, 19.56 and 27.82 % respectively (Fig. 1b). Additionally, the mature renal markers AQP1 and E-cadherin were up-regulated upon RAB stimulation (Fig. 1a). To improve the differentiation efficiency of iPSCs into RPCs, the cells were cultured in renal epithelial cell growth medium for another 5 days (Additional file 1: Figure S1). As expected, the expression of the RPCs markers was further up-regulated compared with that of the RAB group, as shown by real-time PCR analysis after 5 days of differentiation in RAB medium (Fig. 1a). Moreover, the PAX2, WT1, and CD24 single-positive cells increased to 52.56, 27.8 and 36.24 % respectively (Fig. 1b), indicating that more RPCs were generated in the RAB + REGM group.Fig. 1


Transplantation of induced pluripotent stem cell-derived renal stem cells improved acute kidney injury.

Li Q, Tian SF, Guo Y, Niu X, Hu B, Guo SC, Wang NS, Wang Y - Cell Biosci (2015)

Efficient differentiation of renal progenitor cells from mouse induced pluripotent stem cells (iPSCs). The renal differentiation of mouse iPSCs was initiated by the formation of embryoid bodies (EBs). After 2 days, retinoic acid, activin A, and BMP7 were added to the differentiation medium, and the cells were exposed to this supplemented media for 5 days (RAB group). After incubation with growth factors for 5 days, the EBs in the RAB + REGM group were cultured in renal epithelial cell growth medium for another 5 days. a Real-time PCR analysis of the expression of mesoderm (Bry) and renal lineage genes (Pax2, Osr1, WT1, Six2, CD24, Sal1, AQP1, E-cad and PDGFR). b Flow cytometry analysis of the expression of mesoderm renal progenitor-related genes. Black line, cells stained with isotype control; Red line, cells stained with indicated antibodies. *P < 0.05
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4541730&req=5

Fig1: Efficient differentiation of renal progenitor cells from mouse induced pluripotent stem cells (iPSCs). The renal differentiation of mouse iPSCs was initiated by the formation of embryoid bodies (EBs). After 2 days, retinoic acid, activin A, and BMP7 were added to the differentiation medium, and the cells were exposed to this supplemented media for 5 days (RAB group). After incubation with growth factors for 5 days, the EBs in the RAB + REGM group were cultured in renal epithelial cell growth medium for another 5 days. a Real-time PCR analysis of the expression of mesoderm (Bry) and renal lineage genes (Pax2, Osr1, WT1, Six2, CD24, Sal1, AQP1, E-cad and PDGFR). b Flow cytometry analysis of the expression of mesoderm renal progenitor-related genes. Black line, cells stained with isotype control; Red line, cells stained with indicated antibodies. *P < 0.05
Mentions: To investigate whether iPSC-derived RPCs can improve AKI, we first used a nephrogenic cocktail of factors (including retinoic acid, activin A, and BMP7) to induce iPSCs differentiation into RPCs (Additional file 1: Figure S1). The iPS cell line used in this study is tagged with a ubiquitously expressed gfp gene. The real-time PCR analysis showed that the expression of the RPC markers PAX2, WT1, Six2 and CD24 increased after RAB treatment (Fig. 1a), indicating that RPCs were successfully induced. As shown by flow cytometry analysis, the PAX2, WT1 and CD24 positive cells were 33.16, 19.56 and 27.82 % respectively (Fig. 1b). Additionally, the mature renal markers AQP1 and E-cadherin were up-regulated upon RAB stimulation (Fig. 1a). To improve the differentiation efficiency of iPSCs into RPCs, the cells were cultured in renal epithelial cell growth medium for another 5 days (Additional file 1: Figure S1). As expected, the expression of the RPCs markers was further up-regulated compared with that of the RAB group, as shown by real-time PCR analysis after 5 days of differentiation in RAB medium (Fig. 1a). Moreover, the PAX2, WT1, and CD24 single-positive cells increased to 52.56, 27.8 and 36.24 % respectively (Fig. 1b), indicating that more RPCs were generated in the RAB + REGM group.Fig. 1

Bottom Line: However, the therapeutic effect of iPSC-derived RPCs for AKI has yet to be determined.We then established the rat ischemia-reperfusion injury (IR) model and transplanted the iPSC-derived RPCs into the injured rats in combination with the hydrogel.Our results revealed that iPSC-derived RPCs can protect AKI rat from renal function impairment and severe tubular injury by up-regulating the renal tubules formation, promoting cell proliferation, reducing apoptosis, and regulating the microenvironment in the injured kidney.

View Article: PubMed Central - PubMed

Affiliation: Institute of Microsurgery on Extremities, Shanghai Jiao Tong University Affiliated Sixth People's Hospital, Shanghai, 200233 China.

ABSTRACT

Background: Acute kidney injury (AKI) is a severe disease with high morbidity and mortality. Methods that promote repair of the injured kidney have been extensively investigated. Cell-based therapy with mesenchymal stem cells or renal progenitor cells (RPCs) resident in the kidney has appeared to be an effective strategy for the treatment of AKI. Embryonic stem cells or induced pluripotent stem cells (iPSCs) are also utilized for AKI recovery. However, the therapeutic effect of iPSC-derived RPCs for AKI has yet to be determined.

Methods: In this study, we induced iPSCs differentiation into RPCs using a nephrogenic cocktail of factors combined with the renal epithelial cell growth medium. We then established the rat ischemia-reperfusion injury (IR) model and transplanted the iPSC-derived RPCs into the injured rats in combination with the hydrogel. Next, we examined the renal function-related markers and renal histology to assess the therapeutic effect of the injected cells. Moreover, we investigated the mechanism by which iPSC-derived RPCs affect AKI caused by IR.

Results: We showed that the differentiation efficiency of iPSCs to RPCs increased when cultured with renal epithelial cell growth medium after stimulation with a nephrogenic cocktail of factors. The transplantation of iPSC-derived RPCs decreased the levels of biomarkers indicative of renal injury and attenuated the necrosis and apoptosis of renal tissues, but resulted in the up-regulation of renal tubules formation, cell proliferation, and the expression of pro-renal factors.

Conclusion: Our results revealed that iPSC-derived RPCs can protect AKI rat from renal function impairment and severe tubular injury by up-regulating the renal tubules formation, promoting cell proliferation, reducing apoptosis, and regulating the microenvironment in the injured kidney.

No MeSH data available.


Related in: MedlinePlus