Homing Of MSCs Research Articles

Kidney cancer cells secrete IL-8 to activate Akt and promote migration of mesenchymal stem cells

BackgroundMesenchymal stem cells (MSCs) are multipotent adult stem cells that have the capability of homing to cancer cells. Thus, MSCs play an important role in the development, metastasis, and drug resistance of cancers. The mechanisms underlying the homing of MSCs in kidney cancer are still poorly understood. MethodsIn the present study, enzyme-linked immunosorbent assay was used to measure the level of IL-8 in patients with kidney cancer and in the culture medium of kidney cancer cells. Immunofluorescence staining and reverse transcription polymerase chain reaction were utilized to explore the main receptor for IL-8 in MSCs. Transwell migration assay was performed to measure the migration ability of MSCs and Western blot test was performed to test the activation of signaling pathways. ResultsThe serum level of IL-8 was markedly increased in patients with kidney cancer, and 2 kidney cancer cell lines were found to secrete IL-8. MSCs had high expression of the IL-8 receptor (CXCR2). Blocking IL-8 or CXCR2 could decrease the migration ability of MSCs. IL-8 could significantly increase Akt phosphorylation in MSCs. ConclusionsKidney cancer cells secrete IL-8 to activate the Akt signaling pathway via CXCR2 on MSCs, inducing the migration of MSCs, which may be one of the important mechanisms underlying the homing of MSCs in kidney cancer.

Protective Effects of Mesenchymal Stem Cells with CXCR4 Up-Regulation in a Rat Renal Transplantation Model

The homing of mesenchymal stem cells to injured tissue, which is important for the correction of conditions such as ischemia-reperfusion injury (IRI) and immunolesions, has been performed previously, but with poor efficiency. Substantial improvements in engraftment are required to derive clinical benefits from MSC transplantation. Chemokines are the most important factors that control cellular migration. Stromal derived factor-1 (SDF-1) is up-regulated during tissue/organ ischemia damage, and its cognate receptor, chemokine receptor 4 (CXCR4), is involved in stem cell migration. The aim of our study was to investigate CXCR4 expression in MSCs and to validate both its role in mediating migration to transplanted kidneys and its immunoregulatory effects in renal protection. Specifically, the present study was designed to investigate the short-term tissue homing of MSCs carrying genetically modified CXCR4 in a rat renal transplantation model. We tested the hypothesis that MSCs with CXCR4 over-expression can more efficiently regulate immunological reactions. Lentiviral vectors were used to over-express CXCR4 or to introduce a short hairpin ribonucleic acid (shRNA) construct targeting endogenous CXCR4 in rat MSCs. MSCs were labeled with enhanced green fluorescent protein (eGFP). After cell sorting, recipient kidneys were regionally perfused; recipient animals were injected with transduced MSCs, native MSCs, or PBS via tail vein following renal transplantation; and the effects of MSC injection were observed.

Chimerism of allogeneic mesenchymal cells in bone marrow, liver, and spleen after mesenchymal stem cells infusion

Although an infusion of culture-expanded MSCs is applied in clinic to improve results of HSCs transplantation and for a treatment of musculoskeletal disorders, homing, and engraftment potential of culture-expanded MSC in humans is still obscure. We report two female patients who received allogeneic BM transplantation as a treatment of hematological diseases and a transplantation of MSCs from third-party male donors. Both patients died within one yr of infectious complications. Specimens of paraffin-embedded blocks of tissues from transplanted patients were taken. The aim of the study was to estimate possible homing and engraftment of allogeneic BM-derived MSCs in some tissues/organs of recipient. Sensitive real-time quantitative PCR analysis was applied with SRY gene as a target. MSC chimerism was found in BM, liver, and spleen of both patients. We conclude that sensitive RQ-PCR analysis is acceptable for low-level chimerism evaluation even in paraffin-embedded tissue specimens.

The homing of bone marrow MSCs to non-osseous sites for ectopic bone formation induced by osteoinductive calcium phosphate

Osteoinductive biomaterials are promising for bone repair. There is no direct proof that bone marrow mesenchymal stem cells (BMSCs) home to non-osseous sites and participate in ectopic bone formation induced by osteoinductive bioceramics. The objective of this study was to use a sex-mismatched beagle dog model to investigate BMSC homing via blood circulation to participate in ectopic bone formation via osteoinductive biomaterial. BMSCs of male dogs were injected into female femoral marrow cavity. The survival and stable chimerism of donor BMSCs in recipients were confirmed with polymerase chain reaction (PCR) and fluorescence in situ hybridization (FISH). Biphasic calcium phosphate (BCP) granules were implanted in dorsal muscles of female dogs. Y chromosomes were detected in samples harvested from female dogs which had received male BMSCs. At 4 weeks, cells with Y-chromosomes were distributed in the new bone matrix throughout the BCP granule implant. At 6 weeks, cells with Y chromosomes were present in newly mineralized woven bone. TRAP positive osteoclast-like cells were observed in 4-week implants, and the number of such cells decreased from 4 to 6 weeks. These results show that osteoprogenitors were recruited from bone marrow and homed to ectopic site to serve as a cell source for calcium phosphate-induced bone formation. In conclusion, BMSCs were demonstrated to migrate from bone marrow through blood circulation to non-osseous bioceramic implant site to contribute to ectopic bone formation in a canine model. BCP induced new bone in muscles without growth factor delivery, showing excellent osteoinductivity that could be useful for bone tissue engineering.

Kidney-Targeted Transplantation of Mesenchymal Stem Cells by Ultrasound-Targeted Microbubble Destruction Promotes Kidney Repair in Diabetic Nephropathy Rats

We test the hypothesis that ultrasound-targeted microbubble destruction (UTMD) technique increases the renoprotective effect of kidney-targeted transplantation of bone-marrow-derived mesenchymal stem cells (BM-MSCs) in diabetic nephropathy (DN) rats. Diabetes was induced by streptozotocin injection (60 mg/Kg, intraperitoneally) in Sprague-Dawley rats. MSCs were administered alone or in combination with UTMD to DN rats at 4 weeks after diabetes onset. Random blood glucose concentrations were measured at 1, 2, 4, and 8 weeks, and plasma insulin levels, urinary albumin excretion rate (UAER) values, the structures of pancreas and kidney, the expressions of TGF-β1, synaptopodin, and IL-10 were assessed at 8 weeks after MSCs transplantation. MSCs transplantation decreased blood glucose concentrations and attenuated pancreatic islets/β cells damage. The permeability of renal interstitial capillaries and VCAM-1 expression increased after UTMD, which enhanced homing and retention of MSCs to kidneys. MSCs transplantation together with UTMD prevented renal damage and decreased UAER values by inhibiting TGF-β1 expression and upregulating synaptopodin and IL-10 expression. We conclude that MSCs transplantation reverts hyperglycemia; UTMD technique noninvasively increases the homing of MSCs to kidneys and promotes renal repair in DN rats. This noninvasive cell delivery method may be feasible and efficient as a novel approach for personal MSCs therapy to diabetic nephropathy.

Lentiviral-mediated ho-1 gene transduced bone mesenchymal stem cells protects against acute graft-Versus-host disease in vitro and vivo

Abstract 4675 Objective:Bone mesenchymal stem cells (BMSCs) possessing immunoregulatory activities have been evaluated in the treatment of graft-versus-host disease (GVHD). In this study, mice's heme oxygenase 1 (HO-1) was transduced into mice's bone marrow-derived mesenchymal stem cells (mBMSCs), we assessed the immuno-suppressive capacity of lentiviral vector transduced BMSCs expressing HO-1 in BALB/c mice aGVHD model, and the immuno-regulatory effect of mBMSCs on alleviating acute GVHD in vivo was measured to provide laboratory data for gene therapy for aGVHD which used mBMSCs as vehicles. Methods:We cloned mice's HO-1 cDNA from mice's bone marrow and constructed recombinant lentivirus vectors (Lentivirus-V5-D-TOPO-HO-1-EGFP/Lentivirus–V 5-D-TOPO-EGFP) which titer was 1×1011 pfu/mL. These mBMSCs were separated, cultured, purified, and detected by morphology, flow cytometry, osteogenic, adipogenic and chondrogenic induction, and the mRNA level of the neural ganglioside GD2 gene which is a surface marker for the identification of MSCs by RT-PCR. Then recombinant vectors were transferred into mBMSCs, and the expression of EGFP and HO-1 were detected by fluorescence microscope, RT-PCR and Western blot respectively. The immunsuppressive capacity of HO-1 over-expressing mBMSCs was investigated using transwell assay in vitro. And before infusion, the homing of MSC was detected by Flow cytometry. In addition, we established BALB/c mice's aGVHD model after Allo-HSCT, the recombinant vectors tranfected mBMSCs and primary mBMSCs were injected into the BALB/c mice aGVHD model in tail vein, respectively. Four groups were separated in vivo test (Group A: aGVHD control; Group B: aGVHD model injected in mBMSCs; Group C: aGVHD model injected in mBMSCs transfected with EGFP; Group D: aGVHD model injected in mBMSCs transfected with HO-1 gene.) The survival, body weight and clinical score of GVHD in transplanted mice were monitored to evaluate the severity of aGVHD. The aGVHD targeted organ, such as Liver, intestine and lung from mice in each group were obtained for histological examination and pathological score. Plasma concentrations of interleukin (IL)-2, IL-4, IL-6, IL-10, IFN-γ and TNF-γ were also determined using a Cytometric Bead Array. Results:We cloned mice HO-1 gene from mice's bone marrow and constructed the recombinant adenovirus vectors (Lentivirus-V5-D-TOPO-HO-1- EGFP/Lentivirus-V5-D-TOPO-EGFP) successfully. mBMSCs was separated and identified successfully. Fluorescence microscope detected the expression of EGFP, while both RT-PCR and Western blot detected high expression of HO-1 in gene-transfected group cells. It clearly showed that mBMSCs in Group D can migrate more through the polycarbonate filter toward bone marrow micro-environment in the lower chamber than Group A, B, C in vitro, while homing of Group D showed similar trend compared with others groups. In the mice GVHD model, treatment with HO-1 over-expressing BMSCs significantly decreased the mortality rate and attenuated clinical and pathological GVHD scores, and volume of spleen was not more obviously enlarged than others groups. Moreover, compared with control groups, the plasma IL-2, IL-6, IFN-γ and TNF-γ levels in recipients infused with HO-1 over-expressing BMSCs were significantly decreased, while those of IL-4 and IL-10 were increased. Conclusion:Lentiviral vectors carrying the HO-1 were successfully used to transduce mBMSCs. In our report, mBMSCs transferred with lentiviral vectors expressed strongerly immunoregulatory activities to alleviate aGVHD. Not only did HO-1 enhance that migration of mBMSCs, but also strengthen homing of MSCs. In vivo experiments, the evidence of survival rate, body weight, clinical score and pathological score fully proved that HO-1-transduced BMSCs effectively controlled the occurrence of mice's aGVHD following allogeneic BM transplantation, and HO-1 may be a potential target to overcome aGVHD in vivo. Disclosures:Ma:National Natural Science Foundation of China: Research Funding. Li:National Natural Science Foundation of China: Research Funding. Fang:National Natural Science Foundation of China: Research Funding. Chen:National Natural Science Foundation of China: Research Funding. Sun:National Natural Science Foundation of China: Research Funding.

E0074 Effects of transplantation of peripheral blood mesenchymal stem cells with hypoxia preconditioning on postangioplasty restenosis in rabbits

ObjectiveTo investigate the function and the mechanism of transplanting bone marrow derived peripheral blood mesenchymal stem cells (PBMSCs) on restenosis after carotid balloon angioplasty in the model of carotid atherosclerosis...

Differential effect of ischaemic preconditioning on mobilisation and recruitment of haematopoietic and mesenchymal stem cells in porcine myocardial ischaemia-reperfusion

Effects of ischaemic preconditioning (IP) on the mobilisation and recruitment of haematopoietic (HSCs) and mesenchymal stem (MSC) cells were determined in porcine coronary occlusion/reperfusion. Thirty-three pigs underwent percutaneous occlusion of the left anterior descending coronary artery (LAD) for 90 minutes (min), followed by 120 min reperfusion. IP was performed in 16 of the 33 pigs by two cycles of 5 min balloon occlusion/reperfusion prior to the 90 min occlusion (group IP vs. group C). Peripheral blood and myocardial tissue concentration of bone marrow origin HSCs (characterised by coexpression of CD31+, CD90+, CD45+) and MSCs (characterised by coexpression of CD44+, CD90+, CD45-) were measured by flow cytometry in the early phase of IP. Plasma/serum levels of stem cell mobilisation factors (stromal cell-derived factor-1a [SDF-1a], vascular endothelial growth factor [VEGF], tumour necrosis factor a[TNF-a] and interleukin-8 [IL-8]) were measured. IP led to a significant increase in circulating HSCs as compared with the group C (475 +/- 233 vs. 281 +/- 264 /ml, p=0.032) in the early phase of IP. In contrast, a rapid and prolonged decrease in level of circulating MSCs was observed in group IP as compared with group C (19 +/- 12 vs. 32 +/- 17 /ml, p=0.015). The recruitment of HSCs and MSCs in infarct and border zone was significantly greater in IP group, indicating a faster homing of MSCs as compared with the rate of mobilisation. Rapid increase in VEGF, TNF-a and IL-8 levels was induced by IP, which, however, was not correlated with the levels of circulating SCs. In conclusion, IP resulted in differential mobilisation and recruitment of HSCs and MSCs in the early phase of cardioprotection.

Targeting Tumor Stroma Using Engineered Mesenchymal Stem Cells Reduces the Growth of Pancreatic Carcinoma

To analyze the efficacy of engineered mesenchymal stem cell based therapy directed towards pancreatic tumor stroma. Mesenchymal stem cells (MSC) are actively recruited to tumor stroma where they enhance tumor growth and metastases. Upregulation of chemotactic cytokine (CCL5) by MSCs within the tumor stroma has been shown to play a central role in this process. Murine MSCs were engineered to express reporter genes or therapeutic genes under control of the CCL5 promoter and adoptively transferred into mice with growing pancreatic tumors. The effect on tumor growth and metastases was then evaluated. MSCs isolated from bone marrow of C57/Bl6 p53 mice were stably transfected with red fluorescent protein (RFP), enhanced green fluorescent protein (eGFP), or herpes simplex virus (HSV) thymidine kinase (Tk) gene driven by the RANTES promoter. MSCs were intravenously applied once per week over 3 weeks to mice carrying an orthotopic, syngeneic pancreatic Panc02 tumor. eGFP and RFP signals driven by the CCL5 promoter were detected by fluorescence in treated pancreatic tumor samples. The HSV-Tk therapy group treated intraperitoneal with the prodrug ganciclovir 5 to 7 days after stem cell application lead to a 50% reduction of primary pancreatic tumor growth (P < 0.0003, student t test) and reduced liver metastases (0% vs. 60%). The active homing of MSCs into primary pancreatic tumor stroma and activation of the CCL5 promoter was verified using eGFP- and RFP-reporter genes. In the presence of ganciclovir, HSV-Tk transfected MSCs led to a significant reduction of primary pancreatic tumor growth and incidence of metastases.