Use Of Cell Transplantation Research Articles

Transplantation of Embryonic Ventral Mesencephalic Tissue in 6-OHDA Induced Parkinsonism Rat Brain for Cell Based Therapy: A Perspective of Methods

Parkinson’s disease (PD) is characterized as a disease of the basal ganglia, with a progressive degeneration of dopaminergic neurons located in the substantia nigra (SN) and projecting to the striatum with subsequently loss of the nigrostriatal circuit. The potential for therapeutic use of cell transplantation for cell replacement has received a great deal of interest. Transplantation with embryonic ventral mesencephalon (VM) is a therapeutic approach for sporadic form of PD. We established unilaterally 6-OHDA lesioned rat model of Parkinson’s disease. Motor behavioral impairment was found compared with normal rat. Embryonic VM tissue was isolated from E 14 (embryonic 14 days) rat brain. We characterized the VM tissue and dissociated cultured dopaminergic cells with tyrosine hydroxylase (TH) immune staining before transplantation in lesioned brain. We observed that the axons of dopaminergic neurons from transplanted VM graft circles round at the site of transplantation in normal adult rat brain. In this paper, we discuss the detailed methodologies which are very useful in preclinical research of cell based therapies for Parkinson’s disease.

Transplantation of Embryonic Ventral Mesencephalic Tissue in 6-OHDA Induced Parkinsonism Rat Brain for Cell Based Therapy: A Perspective of Methods

Parkinson’s disease (PD) is characterized as a disease of the basal ganglia, with a progressive degeneration of dopaminergic neurons located in the substantia nigra (SN) and projecting to the striatum with subsequently loss of the nigrostriatal circuit. The potential for therapeutic use of cell transplantation for cell replacement has received a great deal of interest. Transplantation with embryonic ventral mesencephalon (VM) is a therapeutic approach for sporadic form of PD. We established unilaterally 6-OHDA lesioned rat model of Parkinson’s disease. Motor behavioral impairment was found compared with normal rat. Embryonic VM tissue was isolated from E 14 (embryonic 14 days) rat brain. We characterized the VM tissue and dissociated cultured dopaminergic cells with tyrosine hydroxylase (TH) immune staining before transplantation in lesioned brain. We observed that the axons of dopaminergic neurons from transplanted VM graft circles round at the site of transplantation in normal adult rat brain. In this paper, we discuss the detailed methodologies which are very useful in preclinical research of cell based therapies for Parkinson’s disease.

The Use of Cell Transplantation in Spinal Cord Injuries

Acute spinal cord injuries are life-changing events that lead to substantial morbidity and mortality, but the role of cell-based treatment for these injuries is unclear. Cell therapy is a rapidly evolving treatment methodology, with basic science and early phase I/II human trials showing promise. Multiple cell lines can be used in cell therapy, including adult or embryonic stem cells, Schwann cells, olfactory ensheathing cells, and induced pluripotent stem cells. Adult stem cells, Schwann cells, and olfactory ensheathing cells are readily available but lack the ability to differentiate into cells of the central nervous system. Mesenchymal stem cells can decrease cell death by modifying the local environment into which they are introduced. Peripheral nerve cells, such as Schwann cells and olfactory ensheathing cells, can myelinate existing axons and foster axonal growth in the central nervous system, and embryonic stem cells can differentiate into neural progenitor stem cells of the central nervous system. Induced pluripotent stem cells are the basis of an emerging technology that has yet to be implemented in human trials but may offer a means of cell therapy without the ethical dilemmas associated with embryonic cells.

Rational Autologous Cell Sources For Therapy of Heart Failure - Vehicles and Targets For Gene and RNA Therapies.

This review focuses on the possibilities for intraoperative processing and isolation of autologous cells, particularly atrial appendage-derived cells (AADCs) and cellular micrografts, and their straightforward use in cell transplantation for heart failure therapy. We review the potential of autologous tissues to serve as sources for cell therapy and consider especially those tissues that are used in surgery but from which the excess is currently discarded as surgical waste. We compare the inculture expanded cells to the freshly isolated ones in terms of evidence-based cost-efficacy and their usability as gene- and RNA therapy vehicles. We also review how financial and authority-based decisions and restrictions sculpt the landscape for patients to participate in academic-based trials. Finally, we provide an insight example into AADCs isolation and processing for epicardial therapy during coronary artery bypass surgery.

The Potential for iPS-Derived Stem Cells as a Therapeutic Strategy for Spinal Cord Injury: Opportunities and Challenges.

Spinal cord injury (SCI) is a devastating trauma causing long-lasting disability. Although advances have occurred in the last decade in the medical, surgical and rehabilitative treatments of SCI, the therapeutic approaches are still not ideal. The use of cell transplantation as a therapeutic strategy for the treatment of SCI is promising, particularly since it can target cell replacement, neuroprotection and regeneration. Cell therapies for treating SCI are limited due to several translational roadblocks, including ethical and practical concerns regarding cell sources. The use of iPSCs has been particularly attractive, since they avoid the ethical and moral concerns that surround other stem cells. Furthermore, various cell types with potential for application in the treatment of SCI can be created from autologous sources using iPSCs. For applications in SCI, the iPSCs can be differentiated into neural precursor cells, neurons, oligodendrocytes, astrocytes, neural crest cells and mesenchymal stromal cells that can act by replacing lost cells or providing environmental support. Some methods, such as direct reprogramming, are being investigated to reduce tumorigenicity and improve reprogramming efficiencies, which have been some of the issues surrounding the use of iPSCs clinically to date. Recently, iPSCs have entered clinical trials for use in age-related macular degeneration, further supporting their promise for translation in other conditions, including SCI.

The ultrastructure of capillaries endotheliocytes in muscular tissue after progenitor fetal liver cells transplantation in patients with chronic limb ischemia

The failure to perform reconstructive-restorative surgical interventions in distal arterial lesions, justifies the necessity to search and develop indirect methods of revascularization. One of the promising methods is the use of cell transplantation to activate the reparation and angiogenesis processes in the affected limb. The results of carried out preclinical studies with modeled ischemia in laboratory animals have shown significant activation of angiogenesis after transplantation of progenitor fetal liver stem cells. Consequently, the cell transplantation has been performed in patients with chronic limb ischemia. Using the electron microscopy techniques it was proved that at the ultrastructural level of endotheliocytes of capillaries in muscular tissue, cell transplantation contributes to substantial activation of angiogenesis in patients with chronic limb ischemia, and is a promising method of indirect revascularization.

Synthetic molecules that protect cells from anoikis and their use in cell transplantation.

One of the major problems encountered in cell transplantation is the low level of survival of transplanted cells due to detachment-induced apoptosis, called anoikis. The present study reports on the chemical synthesis and biological evaluation of water-soluble molecules that protect suspended cells from anoikis. The synthetic molecules bind to and induce clusters of integrins and heparan-sulfate-bound syndecans, two classes of receptors that are important for extracellular matrix-mediated cell survival. Molecular biological analysis indicates that such molecules prolong the survival of suspended NIH3T3 cells, at least in part, by promoting clustering of syndecan-4 and integrin β1 on the cell surface, leading to the activation of small GTPase Rac-1 and Akt. In vivo experiments using animal disease models demonstrated the ability of the molecules to improve cell engraftment. The cluster-inducing molecules may provide a starting point for the design of new synthetic tools for cell-based therapy.

Synthetic Molecules that Protect Cells from Anoikis and Their Use in Cell Transplantation

AbstractOne of the major problems encountered in cell transplantation is the low level of survival of transplanted cells due to detachment‐induced apoptosis, called anoikis. The present study reports on the chemical synthesis and biological evaluation of water‐soluble molecules that protect suspended cells from anoikis. The synthetic molecules bind to and induce clusters of integrins and heparan‐sulfate‐bound syndecans, two classes of receptors that are important for extracellular matrix‐mediated cell survival. Molecular biological analysis indicates that such molecules prolong the survival of suspended NIH3T3 cells, at least in part, by promoting clustering of syndecan‐4 and integrin β1 on the cell surface, leading to the activation of small GTPase Rac‐1 and Akt. In vivo experiments using animal disease models demonstrated the ability of the molecules to improve cell engraftment. The cluster‐inducing molecules may provide a starting point for the design of new synthetic tools for cell‐based therapy.

Stem cells in canine spinal cord injury--promise for regenerative therapy in a large animal model of human disease.

The use of cell transplantation for spinal cord injury is a rapidly evolving field in regenerative medicine. Numerous animal models are currently being used. However, translation to human patients is still a challenging step. Dogs are of increasing importance as a translational model for human disease since there is a greater awareness of the need to increase the quality of preclinical data. The use of dogs ultimately brings benefit to both human and veterinary medicine. In this review we analyze experimental and clinical studies using cell transplantation for canine spinal cord injury. Overall, in experimental studies, transplantation groups showed improvement over control groups. Improvements were measured at the functional, electrophysiological, histological, RNA and protein levels. Most clinical studies support beneficial effects of cell transplantation despite the fact that methodological limitations preclude definitive conclusions. However, the mechanisms of action and underlying the behavior of transplanted cells in the injured spinal cord remain unclear. Overall, we conclude here that stem cell interventions are a promising avenue for the treatment of spinal cord injury. Canines are a promising model that may help bridge the gap between translational research and human clinical trials.

Cat amniotic membrane multipotent cells are nontumorigenic and are safe for use in cell transplantation.

Amnion-derived mesenchymal stem cells (AMSCs) are multipotent cells with an enhanced ability to differentiate into multiple lineages. AMSCs can be acquired through noninvasive methods, and therefore are exempt from the typical ethical issues surrounding stem cell use. The objective of this study was to isolate and characterize AMSCs from a cat amniotic membrane for future application in regenerative medicine. The cat AMSCs were harvested after mechanical and enzymatic digestion of amnion. In culture medium, the cat AMSCs adhered to a plastic culture dish and displayed a fibroblast-like morphology. Immunophenotyping assays were positive for the mesenchymal stem cell-specific markers CD73 and CD90 but not the hematopoietic markers CD34, CD45, and CD79. Under appropriate conditions, the cat AMSCs differentiated into osteogenic, chondrogenic, and adipogenic cell lineages. One advantage of cat AMSCs was nonteratogenicity, assessed 4 weeks post injection of undifferentiated AMSCs into immunodeficient mice. These findings suggest that cat amniotic membranes may be an important and useful source of mesenchymal stem cells for clinical applications, especially for cell or tissue replacement in chronic and degenerative diseases.

Bridging between transplantation therapy and neurotrophic factors in Parkinson's disease.

Parkinson's disease (PD) represents a challenging condition where different therapeutic options have evolved over the course of the last 50 years. The potential for therapeutic use of cell transplantation for cell replacement or for gene delivery of neurotrophic factors has received a great deal of attention. Currently, all available treatment options are directed towards the amelioration of symptoms. A greater understanding of the distinctive pathology underlying PD might offer some novel therapeutic approaches. Transplantation of embryonic ventral mesencephalon (VM) dopaminergic neurons has shown promise in animal studies, but similar transplant procedures have shown limited success in clinical trials. One important issue may be the site of transplantation. Previous studies have transplanted VM into the striatum, which is the target of these neurons. With increased understanding of growth and guidance molecule effecting dopaminergic neurons, it may be feasible to place transplants in the damaged substantia nigra and direct the growth of axons into target regions to reconstruction of midbrain dopamine (DA) circuitry. Our established and on-going understanding of the molecular cues which support directed growth of DA neurons form an important basis for the refinement and optimization of VM grafting procedures, and also the development of new procedures based on the use of stem cells. In this review, we discuss transplantation therapy and how selective guidance molecules could be used to reconstruction of nigrostriatal circuit.

Bridging between transplantation therapy and neurotrophic factors in Parkinson's disease

Parkinson's disease (PD) represents a challenging condition where different therapeutic options have evolved over the course of the last 50 years. The potential for therapeutic use of cell transplantation for cell replacement or for gene delivery of neurotrophic factors has received a great deal of attention. Currently, all available treatment options are directed towards the amelioration of symptoms. A greater understanding of the distinctive pathology underlying PD might offer some novel therapeutic approaches. Transplantation of embryonic ventral mesencephalon (VM) dopaminergic neurons has shown promise in animal studies, but similar transplant procedures have shown limited success in clinical trials. One important issue may be the site of transplantation. Previous studies have transplanted VM into the striatum, which is the target of these neurons. With increased understanding of growth and guidance molecule effecting dopaminergic neurons, it may be feasible to place transplants in the damaged substantia nigra and direct the growth of axons into target regions to reconstruction of midbrain dopamine (DA) circuitry. Our established and on-going understanding of the molecular cues which support directed growth of DA neurons form an important basis for the refinement and optimization of VM grafting procedures, and also the development of new procedures based on the use of stem cells. In this review, we discuss transplantation therapy and how selective guidance molecules could be used to reconstruction of nigrostriatal circuit.

Understanding Myeloma Cancer Stem Cells

Multiple myeloma (MM) is a neoplasm of mature plasma cells [1], characterized by the accumulation of malignant cells in the bone marrow that are dedicated to the production of a monoclonal immunoglobulin [1]. It is also char­ acterized by osteolytic lesions [2], anemia, hyper­ calcemia and renal failure [3]. MM is responsible for approximately 10% of all hematological can­ cers [4] and its incidence is increasing, in part because of an aging trend in the world popula­ tion. Approximately half of all MM patients are older than 70 years and 20% of them are older than 80 years at diagnosis [5]. A major change in the treatment of MM occurred in the last decade, with the discovery of new therapeutic agents, such as immuno­ modulatory drugs (thalidomide and lenalido­ mide) and proteasome inhibitors (bortezomib) [6]. The use of these drugs, in different combi­ nations, improved the complete response rate in patients eligible for autologous hematopoietic stem cell transplantation, and has contributed to prolong progression­free survival and to improve the quality of life of MM patients [7]. However, after more than 30 years of autologous hematopoietic stem cell transplantation use in MM treatment, this procedure is not consid­ ered curative [2]. Even with multiple approaches and promising therapeutic agents, MM remains an incurable disease [3,8]. Thus, a better under­ standing of the molecular pathogenesis of MM is essential for the development of novel drugs capable of overcoming relapses and improving survival in MM.

Clinical Grade Manufacturing of Human Alloantigen-Reactive Regulatory T Cells for Use in Transplantation

Regulatory T cell (Treg) therapy has the potential to induce transplantation tolerance so that immunosuppression and associated morbidity can be minimized. Alloantigen-reactive Tregs (arTregs) are more effective at preventing graft rejection than polyclonally expanded Tregs (PolyTregs) in murine models. We have developed a manufacturing process to expand human arTregs in short-term cultures using good manufacturing practice-compliant reagents. This process uses CD40L-activated allogeneic B cells to selectively expand arTregs followed by polyclonal restimulation to increase yield. Tregs expanded 100- to 1600-fold were highly alloantigen reactive and expressed the phenotype of stable Tregs. The alloantigen-expanded Tregs had a diverse TCR repertoire. They were more potent than PolyTregs in vitro and more effective at controlling allograft injuries in vivo in a humanized mouse model.

Diverse functions of secreted frizzled-related proteins in the osteoblastogenesis of human multipotent mesenchymal stromal cells

Osteoinductive pretreatment of human mesenchymal stromal cells (hMSCs) has been widely accepted in bone tissue engineering before the use of cell transplantation; however, the mechanisms by which osteoinductive medium (OIM) enhances osteoblastic differentiation are not well understood. Using periodontal ligament-derived hMSCs, we identified key signalling molecules for osteoblastogenesis. Alkaline phosphatase activity induced by OIM, which contains ascorbic acid, β-glycerophosphate, and dexamethasone, was decreased by XAV939, which is an inhibitor of canonical WNT signalling, in a dose-dependent manner. A quantitative RT-PCR array demonstrated the upregulation of secreted frizzled-related protein (SFRP) 3 and the downregulation of SFRP4 during osteoinduction. Functional studies showed that SFRP3 promoted and SFRP4 suppressed the osteoblastic differentiation of hMSCs. In addition, SFRP3 inhibited non-canonical WNT signalling by binding WNT5A, which is a representative non-canonical WNT protein. These results indicate the involvement of the WNT signalling pathway during the osteoblastic differentiation of hMSCs. SFRPs oppositely control osteoblastogenesis through canonical and non-canonical pathways and may be useful for directing the lineage of hMSCs in cytotherapeutic use.

Cell transplantation approaches to retinal ganglion cell neuroprotection in glaucoma

Glaucoma is a complex neurodegenerative disease that involves interactions among multiple signaling pathways, ultimately leading to progressive retinal ganglion cell (RGC) death. The development of neuroprotective approaches to glaucoma therapy could preserve vision by modulating these pathologic pathways or by acting directly on RGCs to attenuate cell death and maintain function. Intraocular cell transplantation is being evaluated as one approach to achieve sustained RGC neuroprotection. Unlike traditional pharmacological approaches, transplanted cells might be capable of simultaneously targeting multiple pro-survival pathways via local delivery of secreted factors and/or via modulation of the intraocular microenvironment. Elucidating the mechanisms by which different cell types attenuate RGC death in models of glaucoma may uncover additional novel mechanisms of neuroprotection. In this review, we will discuss the rationale for transplantation-based approaches to neuroprotection for glaucoma and explore the various mechanisms of action proposed to account for RGC neuroprotection achieved by two distinct cell classes that have been studied most extensively for this purpose: glial cells and mesenchymal stem cells.

A method for deriving homogenous population of oligodendrocytes from mouse embryonic stem cells

There is a pressing need for new therapeutics for the generation and transplantation of oligodendrocyte to the white matter to help replace and render injured cells that are lost in demyelinating disease. There are a few protocols describing a homogenous derivation of non-manipulated mouse embryonic stem cells to oligodendrocytes (ES-OL). Moreover, protocols that are successful in producing ES-OL do so with low efficiency. Therefore, we describe clear methodology for differentiation of mouse ES cells to oligodendrocyte to a high degree of homogenity and reproducibility in vitro. In addition, taking advantage of three defined media, we can generate a defined ES to oligodendrocyte lineage while selecting against neurons and astrocytes. More specifically, (1) Glial stem cell defining media (GSCDM), supplemented with appropriate combination of SHH and RA support pro-oligodendrocyte developing neural spheres from ES cells, (2) Oligodendrocyte differentiating media, induces lineage selection of oligodendrocytes progenitors from neural stem cells, and (3) Oligodendrocyte maturation media, supports oligodendrocytes progenitor maturation. Moreover, the ES cell derived oligodendrocytes display mature properites in the prescence of rat dorsal root gangila in vitro. Thus confirming thier potential for use to invesitgate developmental pathways and future potential use of cells in transplantation towards myelin repair.

Amniotic mesenchymal tissue cells inhibit tumor cell line proliferation

Amniotic mesenchymal tissue cells inhibit tumor cell line proliferation

Stem cell-derived neurotrophic support for the neuromuscular junction in spinal muscular atrophy

Importance of the field: Spinal muscular atrophy (SMA) is a neurodegenerative disease characterized by specific degeneration of α-motor neurons in the spinal cord. The use of cell transplantation to restore lost function through cell replacement or prevent further degeneration of motor neurons and synapses through neurotrophic support heralds tremendous hope in the SMA field.Areas covered in this review: Much research has been carried out in the last decade on the use of embryonic stem cells in cell replacement strategies for various neurodegenerative diseases. Cell replacement is contingent on the ability of transplanted cells to integrate and form new functional connections with host cells. In the case of SMA, cell replacement is a tall order in that axons of transplanted cells would be required to grow over long distances from the spinal cord through growth-averse terrain to synapse with muscles in the periphery. The efficacy of neurotrophic support is contingent on the ability of transplanted cells to secrete neurotrophins appropriate for degenerating motor neurons in the spinal cord or development/stability of the neuromuscular junction (NMJ) in the periphery.What the reader will gain: The reader will gain an understanding of the potential of neurotrophins to promote development of the NMJ in a diseased or injured environment.Take home message: Neurotrophins play a major role in NMJ development and thus may be a key factor in the pathogenesis of NMJs in SMA. Further research into the signaling mechanisms involved in NMJ maturation may identify additional mechanisms by which transplanted cells may be of therapeutic benefit.

In Vitro Evaluation of New Possible Cell Engraftment Enhancers for Cell Transplantation

Orthotopic liver transplantation (OLT) is the best treatment to restore liver function in liver failure. The low availability of organs has focused interest on the use of cell transplantation to restore liver function. However, this technique is limited because cells can not bind to liver parenchyma and die soon after perfusion. Pretransplant treatment with engraftment enhancers (EE) to increase vascular permeability may increase cell attachment. Using an endothelial cell culture to measure the loss of intercellular endothelial adhesion as a screening test, we evaluated the capacity of 15 monoclonal antibodies against adhesion molecules expressed on endothelial cells to act as EE showing that 3 antibodies (anti-CD54, efalizumab, and abciximab) act as EE by producing disruptions in the cell layer.