Extracellular vesicles from three dimensional culture of human placental mesenchymal stem cells ameliorated renal ischemia/reperfusion injury

Abstract

Background:

Three-dimensional (3D) culture has been reported to increase the therapeutic potential of mesenchymal stem cells (MSCs). The present study assessed the therapeutic efficacy of extracellular vesicles (EVs) from 3D cultures of human placental MSCs (hPMSCs) for acute kidney injury (AKI).

Methods:

The supernatants from monolayer culture (2D) and 3D culture of hPMSCs were ultra-centrifuged for EVs isolation. C57BL/6 male mice were submitted to 45 min bilateral ischemia of kidney, followed by renal intra-capsular administration of EVs within a 72 h reperfusion period. Histological, immunohistochemical, and ELISA analyses of kidney samples were performed to evaluate cell death and inflammation. Kidney function was evaluated by measuring serum creatinine and urea nitrogen. The miRNA expression profiles of EVs from 2D and 3D culture of hPMSCs were evaluated using miRNA microarray analysis.

Results:

The 3D culture of hPMSCs formed spheroids with different diameters depending on the cell density seeded. The hPMSCs produced significantly more EVs in 3D culture than in 2D culture. More importantly, injection of EVs from 3D culture of hPMSCs into mouse kidney with ischemia-reperfusion (I/R)-AKI was more beneficial in protecting from progression of I/R than those from 2D culture. The EVs from 3D culture of hPMSCs were more efficient against apoptosis and inflammation than those from 2D culture, which resulted in a reduction in tissue damage and amelioration of renal function. MicroRNA profiling analysis revealed that a set of microRNAs were significantly changed in EVs from 3D culture of hPMSCs, especially miR-93-5p.

Conclusion:

The EVs from 3D culture of hPMSCs have therapeutic potential for I/R-AKI.

Keywords

Exosome placental mesenchymal stem cells three-dimensional culture acute kidney injury ischemia/ reperfusion

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References

Pozzoli

Simonini

Manunta

Predicting acute kidney injury: current status and future challenges. J Nephrol 2017; 31: 209–223.

Rewa

Bagshaw

SM.

Acute Kidney injury-epidemiology, outcomes and economics. Nat Rev Nephrol 2014; 10: 193–207.

Renal injury repair: How about the role of stem cells. Adv Exp Med Biol 2019; 1165: 661–670.

Bilic

Zeisberger

Mallik

, et al. Comparative characterization of cultured human term placental epithelial and mesenchymal stromal cells for application in cell therapy. Cell Transplantation 2008; 17(8): 955–968.

Nazarinia

Aboutaleb

Gholamzadeh

, et al. Conditioned medium obtained from human amniotic mesenchymal stem cells attenuates focal cerebral ischemia/ reperfusion injury in rats by targeting mTOR pathway. J Chem Neuroanat 2019; 102: 101707.

Shu

, et al. The beneficial effect of human placental mesenchymal cells in inhibition of inflammation and induction of neuronal repair in EAE mice. J Immunol Res 2018; 24: 5083797.

Zhang

Exosomes derived from mesenchymal stem cells. Int J Mol Sci 2014; 15: 4142–4157.

Lai

Arslan

Lee

, et al. Exosome secreted by MSC reduces myocardial ischemia/reperfusion injury. Stem Cell Res 2010; 4: 214–222.

Théry

Witwer

Aikawa

, et al. Minimal information for studies of extracellular vesicles 2018 (MISEV2018): a position statement of the International Society for Extracellular Vesicles and update of the MISEV2014 guidelines. J Extracell Vesicles 2018; 7(1): 1535750.

10.

Qiu

Zheng

, et al. Mesenchymal stem cell derived extracellular vesicles affect disease outcomes via transfer of microRNAs. Stem Cell Res Ther 2018; 9(1): 320.

11.

Zhu

Pei

Lin

, et al. Exosomes from human-bone marrow derived mesenchymal stem cells protect against renal ischemia/reperfusion injury via transferring miR-199a-3p. J Cell Physiol 2019; 234(12): 23736–23749.

12.

Salomon

Ryan

Sobrevia

, et al. Exosomal signaling during hypoxia mediates microvascular endothelial cell migration and vasculogenesis. PLoS One 2013; 8: 68451.

13.

Kato

Miyaki

Ishitobi

, et al. Exosomes from IL-1b stimulated synovial fibroblasts induce osteoarthritic changes in articular chondrocytes. Arthritis Res Ther 2014; 16: 163.

14.

McKee

Chaudhry

GR.

Advances and challenges in stem cell culture. Colloids Surf B Biointerfaces. 2017; 159: 62–77.

15.

Bartosh

Ylo ̈stalo

Mohammadipoor

, et al. Aggregation of human mesenchymal stromal cells (MSCs) into 3D spheroids enhances their anti-inflammatory properties. Proc Natl Acad Sci USA 2010; 107: 13724–13729.

16.

Ylostalo

Bartosh

Coble

, et al Human mesenchymal stem/stromal cells cultured as spheroids are self-activated to produce prostaglandin E2 that directs stimulated macrophages into an anti-inflammatory phenotype. Stem Cells 2012; 30: 2283–2296.

17.

Kim

Yun

Park

, et al. Three-dimensional spheroid culture increases exosome Secretion from mesenchymal stem cells. Tissue Eng Regen Med 2018; 15(4): 427–436.

18.

Hao

Tong

, et al. LPS-preconditioned mesenchymal stromal cells modify macrophage polarization for resolution of chronic inflammation via exosome-shuttled let-7b. J Transl Med 2015; 13: 308.

19.

Taylor

Peak, Prakash

, et al. Exosomes secreted by placental stem cells selectively inhibit growth of aggressive prostate cancer cells. Biochem Biophys Res Commun 2018; 499(4): 1004–1010.

20.

Gao

Liu

, et al. The use of chitosan-based hydrogel for enhancing the therapeutic benefits of adipose-derived MSCs for acute kidney injury. Biomaterials 2012; 33: 3673–3681.

21.

Zhu

Gongmin

Pei

Lijiao

Lin

Fan

, et al. Exosomes from human-bone-marrow-derived mesenchymal stem cells protect against renal ischemia/reperfusion injury via transferring miR-199a-3p. J Cell Physiol 2019; 234(12): 23736–23749.

22.

Frith

Thomson

Genever

PG.

Dynamic three-dimensional culture methods enhance mesenchymal stem cell properties and increase therapeutic potential. Tissue Eng Part C Methods. 2009; 16: 735–749.

23.

Miceli

Pampalone

Vella

, et al. Comparison of immunosuppressive and angiogenic properties of human placental-derived mesenchymal stem cells between 2D and 3D culture systems. Stem Cells Int 2019; 18:7486279.

24.

Havasi

Borkan

SC.

Apoptosis and acute kidney injury. Kidney Int 2011; 80: 29–40.

25.

Shi

, et al. 3D spheroid culture enhances survival and therapeutic capacities of MSCs injected into ischemic kidney. J Cell Mol Med 2016; 20(7): 1203–1213.

26.

Zhang

Sheng

, et al. Total Coumarins from hydrangea paniculata show renal protective effects in lipopolysaccharide-induced acute kidney injury via anti-inflammatory and antioxidant activities. Front Pharmacol 2017; 14(8): 872.

27.

Bonventre

Yang

Cellular pathophysiology of ischemic acute kidney injury. J Clin Investig 2011; 121: 4210–4221.

28.

Zhang

Liu

, et al. Exosomes/tricalcium phosphate combination scaffolds can enhance bone regeneration by activating the PI3K/Akt signaling pathway. Stem Cell Res Ther 2016; 7(1): 136.

29.

Arslan

Lai

Smeets

, et al. Mesenchymal stem cell-derived exosomes increase ATP levels, decrease oxidative stress and activate PI3K/Akt pathway to enhance myocardial viability and prevent adverse remodeling after myocardial ischemia/reperfusion injury. Stem Cell Res 2013; 10(3): 301–312.

30.

Jang

Rabb

Immune cells in experimental acute kidney injury. Nat Rev Nephrol 2015; 11: 88–101.

31.

Zhou

Jia

, et al. Pharmacological Inhibition of PTEN Aggravates Acute Kidney Injury. Sci Rep 2017; 25: 7(1): 9503.

32.

Chevillet

Kang

Ruf

, et al. Quantitative and stoichiometric analysis of the microRNA content of exosomes. Proc Natl Acad Sci USA 2014; 111: 14888–14893.

33.

Zhao

Zhang

, et al. Genetic communication by extracellular vesicles is an important mechanism underlying stem cell-based therapy-mediated protection against acute kidney injury. Stem Cell Res Ther 2019; 10(1): 119.

34.

Choi

Shin

Bae

IH.

miR-93-5p suppresses cellular senescence by directly targeting Bcl-w and p21. Biochem Biophys Res Commun 2018; 505(4): 1134–1140.

35.

Fabbri

Montagner

Bianchi

, et al. MicroRNA miR-93-5p regulates expression of IL-8 and VEGF in neuroblastoma SK-N-AS cells. Oncol Rep 2016; 35(5): 2866–2872.

36.

Jin

, et al. MiR-93-5p targeting PTEN regulates the NMDA-induced autophagy of retinal ganglion cells via AKT/mTOR pathway in glaucoma. Biomed Pharmacother 2018; 100: 1–7.