Document Type : Original Research Article



Given the functional drawbacks in the efficacy and safety of conventional miRNA delivery approaches, we aimed to assess the fate of mesenchymal stem cells (MSCs) expressing miR-146a-5p into the mice hepatic tissue. MSCs were xeno-transplanted through direct intrahepatic and intraperitoneal routes into immunocompetent and cyclosporine A treated BALB/c mice. DNA extracts from murine organs 24 h and 28 days after xenotransplantation were analyzed by PCR method for detection of human-specific Glyceraldehyde-3-Phosphate Dehydrogenase (GAPDH) sequence. Xeno-transplanted naive MSCs could not be detected after 24 h and 28 days post-infusion at the various organs of mice. Considering the miR-146a-5p expression in MSCs together with negative results of cell engraftment in hepatic tissue, the present study suggests the cell-free secretome-based miR-146a-5p delivery for modulation of altered miRNA expression in hepatic stellate cells.


[1]. Gao B and Radaeva S. Natural killer and natural killer T cells in liver fibrosis. Biochimica et Biophysica Acta (BBA)-Molecular Basis of Disease. 2013; 1832: 1061-69.
[2]. Antaris AL, Robinson JT, Yaghi OK, et al. Ultra-low doses of chirality sorted (6, 5) carbon nanotubes for simultaneous tumor imaging and photothermal therapy. ACS Nano. 2013; 7: 3644-52.
[3]. Kim M-D, Kim S-S, Cha H-Y, et al. Therapeutic effect of hepatocyte growth factor-secreting mesenchymal stem cells in a rat model of liver fibrosis. Experimental & molecular medicine. 2014; 46: 110.
[4]. Miao C-g, Yang Y-y, He X, et al. Wnt signaling in liver fibrosis: progress, challenges and potential directions. Biochimie. 2013; 95: 2326-35.
[5]. Liu B, Li R, Zhang J, et al. MicroRNA-708-3p as a potential therapeutic target via the ADAM17-GATA/STAT3 axis in idiopathic pulmonary fibrosis. Experimental & molecular medicine. 2018; 50: 465.
[6]. Wang Y, Du J, Niu X, et al. MiR-130a-3p attenuates activation and induces apoptosis of hepatic stellate cells in nonalcoholic fibrosing steatohepatitis by directly targeting TGFBR1 and TGFBR2. Cell Death &Amp; Disease. 2017; 8: 2792.
[7]. Caviglia JM, Yan J, Jang MK, et al. MicroRNA‐21 and Dicer are dispensable for hepatic stellate cell activation and the development of liver fibrosis. Hepatology. 2018; 67: 2414-29.
[8]. Wang C, Zhang W, Zhang L, et al. miR-146a-5p mediates epithelial–mesenchymal transition of oesophageal squamous cell carcinoma via targeting Notch2. British J cancer. 2016; 115: 1548.
[9]. Jia XQ, Cheng HQ, Qian X, et al. Lentivirus-mediated overexpression of microRNA-199a inhibits cell proliferation of human hepatocellular carcinoma. Cell biochemistry and biophysics. 2012; 62: 237-44.
[10]. Du J, Niu X, Wang Y, et al. MiR-146a-5p suppresses activation and proliferation of hepatic stellate cells in nonalcoholic fibrosing steatohepatitis through directly targeting Wnt1 and Wnt5a. Sci. Rep. 2015; 5: 16163.
[11]. Ji X, Wu B, Fan J, et al. The anti-fibrotic effects and mechanisms of microRNA-486-5p in pulmonary fibrosis. Sci. Rep. 2015; 5: 14131.
[12]. Jang E, Kim E, Son H-Y, et al. Nanovesicle-mediated systemic delivery of microRNA-34a for CD44 overexpressing gastric cancer stem cell therapy. Biomaterials. 2016; 105: 12-24.
[13]. Fernandez-Piñeiro I, Badiola I and Sanchez A. Nanocarriers for microRNA delivery in cancer medicine. Biotechnology advances. 2017; 35: 350-60.
[14]. Yalcin S and Gunduz U. Nanoparticle based delivery of miRNAs to overcome drug resistance in breast cancer. J. Nanomed. Nanotechnol. 2016; 7: 414.
[15]. Chen Y, Gao D-Y and Huang L. In vivo delivery of miRNAs for cancer therapy: challenges and strategies. Adv. Drug Delivery Rev. 2015; 81: 128-41.
[16]. Collino F, Bruno S, Lindoso RS and Camussi G. miRNA Expression in Mesenchymal Stem Cells. Current Pathobiol Reports. 2014; 2: 101-7.
[17]. Karp JM and Teo GSL. Mesenchymal stem cell homing: the devil is in the details. Cell stem cell. 2009; 4: 206-16.
[18]. Boura JS, Vance M, Yin W, et al. Evaluation of gene delivery strategies to efficiently overexpress functional HLA-G on human bone marrow stromal cells. Molecular Therapy-Methods & Clinical Development. 2014; 1: 14041.
[19]. Amer MH, Rose FR, Shakesheff KM, et al. Translational considerations in injectable cell-based therapeutics for neurological applications: concepts, progress and challenges. NPJ Regenerative medicine. 2017; 2: 23.
[20]. Eggenhofer E, Benseler V, Kroemer A, et al. Mesenchymal stem cells are short-lived and do not migrate beyond the lungs after intravenous infusion. Front. Immunol. 2012; 3: 297.
[21]. Saat TC, van den Engel S, Bijman-Lachger W, et al. Fate and effect of intravenously infused mesenchymal stem cells in a mouse model of hepatic ischemia reperfusion injury and resection. Stem Cells Int. 2016; 2016: 9.
[22]. Fath-Bayati L and Ai J. Assessment of mesenchymal stem cell effect on foreign body response induced by intraperitoneally implanted alginate spheres. J Biomedical Materials Res Part A. 2020; 108: 94-102.
[23]. Lee S-T, Chu K, Kim E-H, et al. Quantification of human neural stem cell engraftments in rat brains using ERV-3 real-time PCR. J Neuroscience Methods. 2006; 157: 225-29.
[24]. Cheng K and Gupta S. Quantitative tools for assessing the fate of xenotransplanted human stem/progenitor cells in chimeric mice. Xenotransplantation. 2009; 16: 145-51.
[25]. Itakura G, Kobayashi Y, Nishimura S, et al. Controlling immune rejection is a fail-safe system against potential tumorigenicity after human iPSC-derived neural stem cell transplantation. PloS one. 2015; 10: 116413.
[26]. Chen Y, Zhao Y, Chen W, et al. MicroRNA-133 overexpression promotes the therapeutic efficacy of mesenchymal stem cells on acute myocardial infarction. Stem Cell Res. Ther. 2017; 8: 268.
[27]. Zhang B, Xuan C, Ji Y, et al. Zebrafish xenotransplantation as a tool for in vivo cancer study. Familial cancer. 2015; 14: 487-93.
[28]. Fischer UM, Harting MT, Jimenez F, et al. Pulmonary passage is a major obstacle for intravenous stem cell delivery: The pulmonary first-pass effect. Stem Cells Dev. 2009; 18: 683-92.
[29]. Heath RD, Ertem F, Romana BS, et al. Hepatocyte transplantation: consider infusion before incision. World J Transplantation. 2017; 7: 317.
[30]. Baccarani U, Adani GL, Sanna A, et al. Portal vein thrombosis after intraportal hepatocytes transplantation in a liver transplant recipient. Transplant International. 2005; 18: 750-54.
[31]. Chamberlain J, Yamagami T, Colletti E, et al. Efficient generation of human hepatocytes by the intrahepatic delivery of clonal human mesenchymal stem cells in fetal sheep. Hepatology. 2007; 46: 1935-45.
[32]. Bazhanov N, Ylostalo JH, Bartosh TJ, et al. Intraperitoneally infused human mesenchymal stem cells form aggregates with mouse immune cells and attach to peritoneal organs. Stem Cell Res. Ther. 2016; 7: 27.
[33]. Wang M, Liang C, Hu H, et al. Intraperitoneal injection (IP), Intravenous injection (IV) or anal injection (AI)? Best way for mesenchymal stem cells transplantation for colitis. Sci. Rep. 2016; 6: 30696.
[34]. Schmuck EG, Koch JM, Centanni JM, et al. Biodistribution and clearance of human mesenchymal stem cells by quantitative three‐dimensional cryo‐imaging after intravenous infusion in a rat lung injury model. Stem Cells Transl. Med. 2016; 5: 1668-75.
[35]. Chuang C-K, Lin K-J, Lin C-Y, et al. Xenotransplantation of human mesenchymal stem cells into immunocompetent rats for calvarial bone repair. Tissue Engineering Part A. 2009; 16: 479-88.
[36]. Talwadekar MD, Kale VP and Limaye LS. Placenta-derived mesenchymal stem cells possess better immunoregulatory properties compared to their cord-derived counterparts–a paired sample study. Sci. Rep. 2015; 5: 15784.
[37]. Fu X, Jiang B, Zheng B, et al. Heterogenic transplantation of bone marrow-derived rhesus macaque mesenchymal stem cells ameliorates liver fibrosis induced by carbon tetrachloride in mouse. PeerJ. 2018; 6: 4336.
[38]. Kim SM, Jeong CH, Woo JS, et al. In vivo near-infrared imaging for the tracking of systemically delivered mesenchymal stem cells: tropism for brain tumors and biodistribution. Int. J. Nanomed. 2016; 11: 13.
[39]. Sun N, Lee A and Wu JC. Long term non-invasive imaging of embryonic stem cells using reporter genes. Nature Protocols. 2009; 4: 1192.
[40]. Becker M, Nitsche A, Neumann C, et al. Sensitive PCR method for the detection and real-time quantification of human cells in xenotransplantation systems. British J Cancer. 2002; 87: 1328.
[41]. Terrovitis JV, Smith RR and Marbán E. Assessment and optimization of cell engraftment after transplantation into the heart. Circ. Res. 2010; 106: 479-94.
[42]. Reyes B, Coca MI, Codinach M, et al. Assessment of biodistribution using mesenchymal stromal cells: Algorithm for study design and challenges in detection methodologies. Cytotherapy. 2017; 19: 1060-9.
[43]. Vrijsen KR, Maring JA, Chamuleau SA, et al. Exosomes from cardiomyocyte progenitor cells and mesenchymal stem cells stimulate angiogenesis via EMMPRIN. Adv Healthcare Materials. 2016; 5: 2555-65.
[44]. Furuta T, Miyaki S, Ishitobi H, et al. Mesenchymal stem cell‐derived exosomes promote fracture healing in a mouse model. Stem Cells Transl. Med. 2016; 5: 1620-30.
[45]. Eirin A, Riester SM, Zhu X-Y, et al. MicroRNA and mRNA cargo of extracellular vesicles from porcine adipose tissue-derived mesenchymal stem cells. Gene. 2014; 551: 55-64.
[46]. Baglio SR, Rooijers K, Koppers-Lalic D, et al. Human bone marrow-and adipose-mesenchymal stem cells secrete exosomes enriched in distinctive miRNA and tRNA species. Stem Cell Res. Ther. 2015; 6: 127.