Transplantation Of Human Mesenchymal Stem Cells Research Articles

Cardiac cell therapy with mesenchymal stem cell induces cardiac nerve sprouting, angiogenesis, and reduced connexin43-positive gap junctions, but concomitant electrical pacing increases connexin43-positive gap junctions in canine heart

Although electrical pacing is of great utility in many cardiovascular diseases, its effects on the combined cardiac cell therapy have not been established. We hypothesised that mesenchymal stem cell transplantation changes cardiac sympathetic nerve and gap junction, and concomitant pacing has additional biological effects. We monitored cardiac rhythm for 4 weeks after human mesenchymal stem cell transplantation (1 x 107, epicardial injection) in 18 dogs in vivo, seven human mesenchymal stem cell with pacing, six human mesenchymal stem cell, and five sham, and evaluated the sympathetic innervation, nerve growth factor-beta; tyrosine hydroxylase, angiogenesis, von Willebrand factor, and connexin43 expressions by real time (RT)-polymerase chain reaction and immunostaining. We also measured mRNA expressions of nerve growth factor-beta, von Willebrand factor, and connexin43 in vitro culture of human mesenchymal stem cell with or without pacing. Human mesenchymal stem cell transplanted hearts expressed higher mRNA of nerve growth factor-beta (p < 0.01) with sympathetic nerves (p < 0.05), higher mRNA of von Willebrand factor (p < 0.001) with angiogenesis (p < 0.001), but lower mRNA of connexin43 (p < 0.0001) with reduced gap junctions (p < 0.001) than sham. Pacing with human mesenchymal stem cell transplantation resulted in higher expression of mRNA of connexin43 (p < 0.02) and gap junctions (p < 0.001) compared with sham. In contrast, in vitro paced mesenchymal stem cell reduced expression of connexin43 mRNA (p < 0.02). Human mesenchymal stem cell transplantation increased cardiac sympathetic innervation and angiogenesis, but reduced gap junction after transplanted in the canine heart. In contrast, concomitant electrical pacing increased gap junction expression by paracrine action.

Human mesenchymal stem cell transplantation protects against cerebral ischemic injury and upregulates interleukin-10 expression in Macacafascicularis

Mesenchymal stem cell (MSC) transplantation has been reported to improve neurologic function after ischemic injury. However, the detailed mechanisms by which MSCs promote functional recovery are not fully understood. Interleukin-10 (IL-10) is a well-known anti-inflammatory cytokine with neuroprotective effects with respect to brain injury. In this study, a non-human primate ischemia model was used to test the hypothesis that transplanted human bone-marrow-derived MSCs (hBMSCs) exert a neuroprotective effects on cerebral ischemia and upregulate IL-10 expression. We also assessed neuronal apoptosis and astroglial activity in the area around the ischemic lesion and proliferating cells in the subventricular zone (SVZ). Results showed that hBMSC transplantation in ischemic tissues improved the neurological functions and induced an increase in IL-10 expression. In addition, neuronal apoptosis and astroglial activity in the peri-ischemic area decreased, and the number of proliferating cells in the SVZ increased. These results provide a novel therapeutic strategy for improving neurologic function after cerebral ischemia.

Xenogenic Transplantation of Human Mesenchymal Stem Cells in a Critical Size Defect of the Sheep Tibia for Bone Regeneration

Mesenchymal stem cells (MSCs) represent an attractive cell population for the regeneration of mesenchymal tissues. Their special immunological characteristics suggest that MSCs could be used in nonautologous applications. In this study, the regenerative capacity of human and ovine MSCs was assessed in an ovine critical size defect model. Human and ovine MSCs from bone marrow were cultured on mineralized collagen and implanted into a 3.0-cm-long sheep tibia bone defect (n = 7). Unloaded mineralized collagen served as control. Bone healing was assessed until euthanasia 26 weeks after surgery by radiology and histologically after euthanasia. The presence of human cells after xenogenic transplantation was analyzed using human-specific in situ hybridization. Both radiology and histology demonstrated significantly better bone formation after transplantation of autologous ovine MSCs on mineralized collagen compared with unloaded matrices and with the xenogenic treatment group. Nevertheless, no local or systemic rejection reactions could be observed after transplantation of human MSCs, although the presence of human MSCs could be demonstrated. It can be concluded that despite successful demonstration of the presence of human MSCs after xenogenic transplantation, the xenogenic transplantation of human MSCs leads to poorer bone regeneration than autologous transplantation of ovine MSCs.

Promotion of incisional wound repair by human mesenchymal stem cell transplantation

The purpose of this study was to determine the effect of transplanted human mesenchymal stem cells (hMSCs) on wound healing. In this model, full-thickness cutaneous wounds were created by incision in the skin of adult New Zealand white rabbits and treated by transplanted hMSCs into the wounds. Wound healing was evaluated by histological analysis and tensiometry over time. A total of 15 New Zealand white rabbits with 10 wounds per animal were examined in this study. Animals were treated with hMSCs and euthanised at 3, 7, 14, 21 and 80 days after manipulation. The hMSCs were labelled with a fluorescent dye (CM-DiI), suspended in phosphate-buffered saline and used to treat full-thickness incisional wounds in rabbit skin. Tensiometry and histology were used to characterise the wound-healing rate of the incisional wounds. These results showed that transplanted hMSCs significantly inhibited scar formation and increased the tensile strength of the wounds. Importantly, MSCs from genetically unrelated donors did not appear to induce an immunologic response. In conclusion, human mesenchymal stem cell therapy is a viable approach to significantly affect the course of normal cutaneous wound healing and significantly increase the tensile strength.

Transplantation of Human Bone Marrow Mesenchymal Stem Cell Ameliorates the Autoimmune Pathogenesis in MRL/lpr Mice

Recent evidence indicates that mesenchymal stem cells (MSC) possess immunosuppressive properties both in vitro and in vivo. We previously demonstrated the functional abnormality of bone marrow derived MSC in patients with systemic lupus erythematosus (SLE). In this study, we aimed to investigate whether transplantation of human bone marrow derived MSC affects the autoimmune pathogenesis in MRL/lpr mice. We found that human MSC from healthy donors reduced the proliferation of T lymphocytes from MRL/lpr mice in a dose-dependent fashion. Two weeks after in vivo transfer of MSC, we detected significantly reduced serum levels of anti ds-DNA antibodies and 24 hour proteinuria in MRL/lpr mice as compared with control groups without MSC transplantation. Moreover, flow cytometric analysis revealed markedly reduced number of CD4(+) T cells while increased Th1 subpopulation in MSC group and MSC + CTX group when compared with controls. Histopathological examination showed significantly reduced renal pathology in MSC-treated mice. Immunohistochemical studies further revealed reduced expression of TGF-beta, FN, VEGF and the deposition of complement C3 in renal tissue after MSC and MSC + CTX treatment. Taken together, we have demonstrated that transplantation of human MSC can significantly inhibit the autoimmune progression in MRL/lpr mice.

Generation of tissue-specific cells from MSC does not require fusion or donor-to-host mitochondrial/membrane transfer

Human mesenchymal stem cells (MSC) hold great promise for cellular replacement therapies. Despite their contributing to phenotypically distinct cells in multiple tissues, controversy remains regarding whether the phenotype switch results from a true differentiation process. Here, we studied the events occurring during the first 120 h after human MSC transplantation into a large animal model. We demonstrate that MSC, shortly after engrafting different tissues, undergo proliferation and rapidly initiate the differentiative process, changing their phenotype into tissue-specific cells. Thus, the final level of tissue-specific cell contribution is not determined solely by the initial level of engraftment of the MSC within that organ, but rather by the proliferative capability of the ensuing tissue-specific cells into which the MSC rapidly differentiate. Furthermore, we show that true differentiation, and not cell fusion or transfer of mitochondria or membrane-derived vesicles between transplanted and resident cells, is the primary mechanism contributing to the change of phenotype of MSC upon transplantation.

Human mesenchymal stem cell transplantation extends survival, improves motor performance and decreases neuroinflammation in mouse model of amyotrophic lateral sclerosis

Amyotrophic lateral sclerosis (ALS) is a lethal disease affecting motoneurons. In familial ALS, patients bear mutations in the superoxide dismutase gene (SOD1). We transplanted human bone marrow mesenchymal stem cells (hMSCs) into the lumbar spinal cord of asymptomatic SOD1 G93A mice, an experimental model of ALS. hMSCs were found in the spinal cord 10 weeks after, sometimes close to motoneurons and were rarely GFAP- or MAP2-positive. In females, where progression is slower than in males, astrogliosis and microglial activation were reduced and motoneuron counts with the optical fractionator were higher following transplantation. Motor tests (Rotarod, Paw Grip Endurance, neurological examination) were significantly improved in transplanted males. Therefore hMSCs are a good candidate for ALS cell therapy: they can survive and migrate after transplantation in the lumbar spinal cord, where they prevent astrogliosis and microglial activation and delay ALS-related decrease in the number of motoneurons, thus resulting in amelioration of the motor performance.

Stem Cell Therapy for Liver Disease: Parameters Governing the Success of Using Bone Marrow Mesenchymal Stem Cells

Liver transplantation is the primary treatment for various end-stage hepatic diseases but is hindered by the lack of donor organs and by complications associated with rejection and immunosuppression. There is increasing evidence to suggest the bone marrow is a transplantable source of hepatic progenitors. We previously reported that multipotent bone marrow-derived mesenchymal stem cells differentiate into functional hepatocyte-like cells with almost 100% induction frequency under defined conditions, suggesting the potential for clinical applications. The aim of this study was to critically analyze the various parameters governing the success of bone marrow-derived mesenchymal stem cell-based therapy for treatment of liver diseases. Lethal fulminant hepatic failure in nonobese diabetic severe combined immunodeficient mice was induced by carbon tetrachloride gavage. Mesenchymal stem cell-derived hepatocytes and mesenchymal stem cells were then intrasplenically or intravenously transplanted at different doses. Both mesenchymal stem cell-derived hepatocytes and mesenchymal stem cells, transplanted by either intrasplenic or intravenous route, engrafted recipient liver, differentiated into functional hepatocytes, and rescued liver failure. Intravenous transplantation was more effective in rescuing liver failure than intrasplenic transplantation. Moreover, mesenchymal stem cells were more resistant to reactive oxygen species in vitro, reduced oxidative stress in recipient mice, and accelerated repopulation of hepatocytes after liver damage, suggesting a possible role for paracrine effects. Bone marrow-derived mesenchymal stem cells can effectively rescue experimental liver failure and contribute to liver regeneration and offer a potentially alternative therapy to organ transplantation for treatment of liver diseases.

Human Mesenchymal Stem Cell Transplantation Induces Sympathetic Nerve Sprouting and Reduces the Gap Junction With Potential Proarrhythmias in Dogs

Background and Objectives: Although human mesenchymal stem cell (hMSC) transplantation has been known to improve ventricular function, the potential proarrhythmic effects have not yet been studied. Materials and Methods: We monitored the heart rhythm of 6 dogs for 4 weeks after transplantation of hMSC (1×10 7 , epicardial injection) (hMSC group) and in 5 Sham dogs after the injection of the vehicle alone. Cardiac sympathetic nerve sprouting {nerve growth factor (NGF)-β; tyrosine hydroxylase (TH)} and gap junction expression {connexin (Cx) 43} were evaluated in 10 dogs (5 hMSC and 5 Sham) that survived longer than 4 weeks. Results: The hMSC group expressed higher levels of NGF-β messenger ribonucleic acid (mRNA) (56.0±66.8 fold; p<0.01) with TH+ sympathetic nerves (0.51±0.40 vs. 0.15±0.13% area; p<0.03) than the Sham control. In contrast, the hMSC group expressed lower levels of Cx43 mRNA (0.59±0.29 fold, p<0.001) and Cx43+ (1.64±1.79 vs. 2.12±1.07% area, p<0.001) than the Sham control. The incidences of ventricular fibrillation were 33.3% and 0% in the hMSC group and Sham control, respectively. One of the dogs with ventricular fibrillation (VF) in the hMSC group died suddenly. Conclusion: hMSC transplantation may be proarrhythmic since NGF-β expression increased with cardiac sympathetic hyperinnervation and the expression of Cx43 and the gap junction decreased. (Korean Circ J 2008;38:536-543)

Human mesenchymal stem cell transplantation promotes functional recovery following acute spinal cord injury in rats

Many attempts have been made in animals to produce cellular regeneration in the spinal cord using a variety of transplanted cell types. The present study was to investigate whether transplantation of human mesenchymal stem cells (hMSCs) into the spinal cord after contusion injury promotes a functional outcome. Spinal cord injury (SCI) was induced using an NYU impactor and hMSCs were transplanted 1 week after SCI. Behavioral testing was performed weekly for 2 months. Somatosensory (SSEPs) and motor evoked potentials (MEPs) were recorded to determine functional recovery. Hindlimb performance was modestly improved in the transplanted group based on BBB scaling and pain tests. SSEP latencies in the transplanted group were significantly shorter than in the media-treated group. Pathologically, LacZ and hTau positive cells were located at the injury and adjacent sites. The data indicate improvement in functional outcome in animals treated with hMSC transplantation compared to media-treated animals. Correspondence should be addressed to B.H. Lee, Email: bhlee@yumc.yonsei.ac.kr

Dynamic of distribution of human bone marrow-derived mesenchymal stem cells after transplantation into adult unconditioned mice.

The use of mesenchymal stem cells (MSC) for cell therapy relies on their capacity to engraft and survive long-term in the appropriate target tissue(s). Animal models have demonstrated that the syngeneic or xenogeneic transplantation of MSC results in donor engraftment into the bone marrow and other tissues of conditioned recipients. However, there are no reliable data showing the fate of human MSC infused into conditioned or unconditioned adult recipients. In the present study, the authors investigated, by using imaging, polymerase chain reaction (PCR), and in situ hybridization, the biodistribution of human bone marrow-derived MSC after intravenous infusion into unconditioned adult nude mice. As assessed by imaging (gamma camera), PCR, and in situ hybridization analysis, the authors' results demonstrate the presence of human MSC in bone marrow, spleen, and mesenchymal tissues of recipient mice. These results suggest that human MSC transplantation into unconditioned recipients represents an option for providing cellular therapy and avoids the complications associated with drugs or radiation conditioning.