Development Of Stem Cell Technology Research Articles

Autologous Pluripotent Stem Cell-Derived β-Like Cells for Diabetes Cellular Therapy.

Development of stem cell technologies for cell replacement therapy has progressed rapidly in recent years. Diabetes has long been seen as one of the first applications for stem cell-derived cells because of the loss of only a single cell type-the insulin-producing β-cell. Recent reports have detailed strategies that overcome prior hurdles to generate functional β-like cells from human pluripotent stem cells in vitro, including from human induced pluripotent stem cells (hiPSCs). Even with this accomplishment, addressing immunological barriers to transplantation remains a major challenge for the field. The development of clinically relevant hiPSC derivation methods from patients and demonstration that these cells can be differentiated into β-like cells presents a new opportunity to treat diabetes without immunosuppression or immunoprotective encapsulation or with only targeted protection from autoimmunity. This review focuses on the current status in generating and transplanting autologous β-cells for diabetes cell therapy, highlighting the unique advantages and challenges of this approach.

The application of Mir-183 family and mesenchymal stem cells: A possibility for restoring hearing loss

Hearing loss as one of the most common disabilities approximately over 5% of the world's population – 360 million people – has disabling hearing loss (328 million adults and 32 million children). Recent developments in stem cell technology provide new opportunities for the treatment of deafness. miRNAs are essential factors of an extensively conserved posttranscriptional process controlling gene expression at mRNA level. Various biological processes such as growth and differentiation are regulated by miRNAs. In this review paper we have discussed about the application of miR-183 family and mesenchymal stem cells as a possibility for restoring hearing loss. In this regards, the web of Science and PubMed databases were searched using the Endnote software for the publications about the application of miR-183 family and mesenchymal stem cells (MSCs) to study hearing loss published from 2000 to 2016. The miR-183 family (miR-183, miR-96, and miR-182) is expressed abundantly in sensory cells in inner ear. miR-183 family is significant for the development and persistence of auditory neurons and hair cell. These four genes, i.e. Sox2, Notch1, Jag1, and Hes1, are potentially the targets of miR-183 family. In studies on animal models such as mouse and zebrafish, the time of Atoh1 expression in the hair cells was found to be the E12/5-E14/5 day, and miR-183 family was reported to begin to express on the E14/5 day. Use of human MSCs in differentiating into hair cells has been investigated, demonstrating that MSCs have neuroregenerative capacity. Cell therapy-targeting regeneration of the auditory neurons and hair cell may therefore be a powerful strategy to cure hearing loss that cannot be reversed by current therapies. A combination of the MSCs, specific growth factors and miR-183 cluster (96-182-183) can increase the potential to differentiate into the auditory neurons and hair cell.

Intermittent high oxygen influences the formation of neural retinal tissue from human embryonic stem cells.

The vertebrate retina is a highly multilayered nervous tissue with a large diversity of cellular components. With the development of stem cell technologies, human retinas can be generated in three-dimensional (3-D) culture in vitro. However, understanding the factors modulating key productive processes and the way that they influence development are far from clear. Oxygen, as the most essential element participating in metabolism, is a critical factor regulating organic development. In this study, using 3-D culture of human stem cells, we examined the effect of intermittent high oxygen treatment (40% O2) on the formation and cellular behavior of neural retinas (NR) in the embryonic body (EB). The volume of EB and number of proliferating cells increased significantly under 40% O2 on day 38, 50, and 62. Additionally, the ratio of PAX6+ cells within NR was significantly increased. The neural rosettes could only develop with correct apical-basal polarity under 40% O2. In addition, the generation, migration and maturation of retinal ganglion cells were enhanced under 40% O2. All of these results illustrated that 40% O2 strengthened the formation of NR in EB with characteristics similar to the in vivo state, suggesting that the hyperoxic state facilitated the retinal development in vitro.

A Stem Cell-Derived Platform for Studying Single Synaptic Vesicles in Dopaminergic Synapses.

The exocytotic release of dopamine is one of the most characteristic but also one of the least appreciated processes in dopaminergic neurotransmission. Fluorescence imaging has yielded rich information about the properties of synaptic vesicles and the release of neurotransmitters in excitatory and inhibitory neurons. In contrast, imaging-based studies for in-depth understanding of synaptic vesicle behavior in dopamine neurons are lagging largely because of a lack of suitable preparations. Midbrain culture has been one of the most valuable preparations for the subcellular investigation of dopaminergic transmission; however, the paucity and fragility of cultured dopaminergic neurons limits their use for live cell imaging. Recent developments in stem cell technology have led to the successful production of dopamine neurons from embryonic or induced pluripotent stem cells. Although the dopaminergic identity of these stem cell-derived neurons has been characterized in different ways, vesicle-mediated dopamine release from their axonal terminals has been barely assessed. We report a more efficient procedure to reliably generate dopamine neurons from embryonic stem cells, and it yields more dopamine neurons with more dopaminergic axon projections than midbrain culture does. Using a collection of functional measurements, we show that stem cell-derived dopamine neurons are indistinguishable from those in midbrain culture. Taking advantage of this new preparation, we simultaneously tracked the turnover of hundreds of synaptic vesicles individually using pH-sensitive quantum dots. By doing so, we revealed distinct fusion kinetics of the dopamine-secreting vesicles, which is consistent within both preparations.

The Potential of Stem Cells for the Restoration of Auditory Function in Humans

Hearing loss is one of the most common disabilities, affecting approximately 10% of the population. Hair cells and spiral ganglion neurons are usually damaged in most cases of hearing loss. Currently, there is virtually no biological approach to replace damaged hearing cells. Recent developments in stem cell technology provide new opportunities for the treatment of deafness. Two major strategies have been investigated: differentiation of endogenous stem cells into new hair cells; and introduction of exogenous cells into the inner ear to substitute injured hearing neurons. Although there is still a learning curve in stem cell-based replacement, the probability exists to utilize personalized stem cells to eventually provide a novel intervention for patients with deafness in future clinical research trials.

Stem Cell Therapy in the Philippines a Changing Landscape

The Department of Health recently released guidelines on Stem Cell and Cell-based Therapy known as the Administrative Order 2013-0012, Rules and Regulations Governing the Accreditation of Health Facilities Engaging in Human Stem Cell and Cell-based or Cellular Therapies in the Philippines. This guideline was motivated by the massive proliferation of clinics or laboratories taking advantage of vulnerable uninitiated patients. If unabated, this unregulated procedure may harm people and impact the development of stem cell technology. Basic queries that will guide a prospective stem cell patient will be presented. To date, four hospitals are recognized by the Department of Health to practice stem-cell therapy. These hospitals are Makati Medical Center, The Medical City, National Kidney and Transplant Institute, and the Lung Center of the Philippines. This communication will focus on the current laboratory safety and monitoring practices carried out at the Cellular Therapeutics Laboratory of Makati Medical Center. The Cellular Therapeutics Laboratory is classified as ISO 14644-1 clean room ISO 5 class facility. The presentation will also feature laboratory standards such as autologous Cell Transplant release criteria and critical quality control tests. The future of Stem Cell Therapy will be profoundly influenced by the clinicians, scientists, regulatory offices, media, and society, who are in position to distinguish the diference between science and fiction. Patients who are left with limited clinical options are often entangled; wading through the investigative and controversial scientific breakthroughs is a tight walk.

Spermatogonial Stem Cells (SSCs) in Buffalo (Bubalus bubalis) Testis

BackgroundWater buffalo is an economically important livestock species and about half of its total world population exists in India. Development of stem cell technology in buffalo can find application in targeted genetic modification of this species. Testis has emerged as a source of pluripotent stem cells in mice and human; however, not much information is available in buffalo.Objectives and MethodsPou5f1 (Oct 3/4) is a transcription factor expressed by pluripotent stem cells. Therefore, in the present study, expression of POU5F1 transcript and protein was examined in testes of both young and adult buffaloes by semi-quantitative reverse transcriptase-polymerase chain reaction (RT-PCR) and immunohistochemical analysis. Further, using the testis transplantation assay, a functional assay for spermatogonial stem cells (SSCs), stem cell potential of gonocytes/spermatogonia isolated from prepubertal buffalo testis was also determined.ResultsExpression of POU5F1transcript and protein was detected in prepubertal and adult buffalo testes. Western blot analysis revealed that the POU5F1 protein in the buffalo testis exists in two isoforms; large (∼47 kDa) and small (∼21 kDa). Immunohistochemical analysis revealed that POU5F1 expression in prepubertal buffalo testis was present in gonocytes/spermatogonia and absent from somatic cells. In the adult testis, POU5F1 expression was present primarily in post-meiotic germ cells such as round spermatids, weakly in spermatogonia and spermatocytes, and absent from elongated spermatids. POU5F1 protein expression was seen both in cytoplasm and nuclei of the stained germ cells. Stem cell potential of prepubertal buffalo gonocytes/spermatogonia was confirmed by the presence of colonized DBA-stained cells in the basal membrane of seminiferous tubules of xenotransplanted mice testis.Conclusion/SignificanceThese findings strongly indicate that gonocytes/spermatogonia, isolated for prepubertal buffalo testis can be a potential target for establishing a germ stem cell line that would enable genetic modification of buffaloes.

Progress on stem cell research towards the treatment of Parkinson's disease

Parkinson's disease (PD) is a neurodegenerative disorder characterized by the progressive accumulation of Lewy body inclusions along with selective destruction of dopaminergic (DA) neurons in the nigrostriatal tract of the brain. Genetic studies have revealed much about the pathophysiology of PD, enabling the identification of both biomarkers for diagnosis and genetic targets for therapeutic treatment, which are evolved in tandem with the development of stem cell technologies. The discovery of induced pluripotent stem (iPS) cells facilitates the derivation of stem cells from adult somatic cells for personalized treatment and thus overcomes not only the limited availability of human embryonic stem cells but also ethical concerns surrounding their use. Non-viral, non-integration, or non-DNA-mediated reprogramming technologies are being developed. Protocols for generating midbrain DA neurons are undergoing constant refinement. The iPS cell-derived DA neurons provide cellular models for investigating disease progression in vitro and for screening molecules of novel therapeutic potential and have beneficial effects on improving the behavior of parkinsonian animals. Further progress in the development of safer non-viral/non-biased reprogramming strategies and the subsequent generation of homogenous midbrain DA neurons shall pave the way for clinical trials. A combined approach of drugs, cell replacement, and gene therapy to stop disease progression and to improve treatment may soon be within our reach.

Parkinson's UK: Pushing the Search for a Cure to New Levels

Every hour, someone in the UK is told they have Parkinson’s disease. Becausewe are here, no one has to face Parkinson’s alone. We bring people with Parkinson’s, their caregivers, and their families together via our network of local groups, our website, and our free confidential helpline. Specialist nurses, our supporters, and staff provide information and training on every aspect of Parkinson’s. As the UK’s Parkinson’s support and research charity, we are leading the work to find a cure, andwe are closer than ever.We also campaign to change attitudes and demand better services. Parkinson’s UK’s fight against the disease began in 1969 when Mali Jenkins founded the Parkinson’s Disease Society. Her sister, Sarah Jenkins, had been living with Parkinson’s for some years and was cared for at the family home. Mali looked for patient associations to help people affected by Parkinson’s, but searches of local libraries showed that no such thing existed. Frustrated to find no literature in layman’s terms about her sister’s condition, Mali decided to find outwhat she could about Parkinson’s. She placed advertisements in the personal columns of a couple of newspapers asking people to come forward who were interested in forming an exploratory steering group. Mali chaired the committee, and her sister, Eryl, was secretary. Other friends and family were asked to help. The first meeting took place at Westminster Hospital with just 16 people in attendance. But that event spawned the idea for a national charity dedicated to helping Parkinson’s patients and their caregivers—the Parkinson’s Disease Society, which officially began on February 26, 1969. Today, the organization has grown to include 350 groups spread across the UK dedicated to offering friendship and support to everyone affected by Parkinson’s. In 2010 we changed our name to Parkinson’s UK to reflect our expanded mission, which includes funding research aimed at finding a cure. Alongwith being at the forefront of developing induced pluripotent stem (iPS) cells, Parkinson’s UK has played a key role in campaigning for the implementation of legislation to allow stem cell research to be carried out within the UK, including the 2001 amendment by the British Parliament to the Human Fertilisation and Embryology Act 1990. We also campaigned in 2009 for legislation to allow for the use of cytoplasmic hybrid embryonic stem cells.We aremembers of the UK Stem Cell Funders Forum, which monitors developments in stem cell technology and ensures that the legislation is appropriate for these technologies to develop.

Using induced pluripotent stem cells (iPSC) to model human neuromuscular connectivity: promise or reality?

Motor neuron diseases (MND) such as amyotrophic lateral sclerosis and spinal muscular atrophy are devastating, progressive and ultimately fatal diseases for which there are no effective treatments. Recent evidence from systematic studies of animal models and human patients suggests that the neuromuscular junction (NMJ) is an important early target in MND, demonstrating functional and structural abnormalities in advance of pathological changes occurring in the motor neuron cell body. The ability to study pathological changes occurring at the NMJ in humans is therefore likely to be important for furthering our understanding of disease pathogenesis, and also for designing and testing new therapeutics. However, there are many practical and technical reasons why it is not possible to visualise or record from NMJs in pre- and early-symptomatic MND patients in vivo. Other approaches are therefore required. The development of stem cell technologies has opened up the possibility of creating human NMJs in vitro, using pluripotent cells generated from healthy individuals and patients with MND. This review covers historical attempts to develop mature and functional NMJs in vitro, using co-cultures of muscle and nerve from animals, and discusses how recent developments in the generation and specification of human induced pluripotent stem cells provides an opportunity to build on these previous successes to recapitulate human neuromuscular connectivity in vitro.

Promises of stem cell therapy for retinal degenerative diseases

With the development of stem cell technology, stem cell-based therapy for retinal degeneration has been proposed to restore the visual function. Many animal studies and some clinical trials have shown encouraging results of stem cell-based therapy in retinal degenerative diseases. While stem cell-based therapy is a promising strategy to replace damaged retinal cells and ultimately cure retinal degeneration, there are several important challenges which need to be overcome before stem cell technology can be applied widely in clinical settings. In this review, different types of donor cell origins used in retinal treatments, potential target cell types for therapy, methods of stem cell delivery to the eye, assessments of potential risks in stem cell therapy, as well as future developments of retinal stem cells therapy, will be discussed.

The Impact of Legislative Framework Conditions on the Development of Stem Cell Technology: Assessment of National Innovation Systems

Abstract Stem cells have the potential to launch a new area of medicine, curing deadly diseases with custom-made tissues and organs. But science and technology may have to take note of regulatory developments, which might impact if, how, and where that hope will be realized. Until now, no data have been available on the influence of national human stem cell research legislation on the technological activity in the field of stem cells. In this contribution, we assess whether legislative framework conditions affect technological activity in this field. Stem cell patent activity (US Patent and Trademark Office [USPTO]/European Patent Office [EPO]) and legislative framework conditions with respect to human stem cell research have been identified and characterized for the period 1997–2003 for 23 countries. For these countries, differences in human stem cell legislation coincide with differences in technological productivity. Where obtaining human stem cells from supernumerary embryos is permitted, higher level...

Stem cells in brain diseases: From cell replacement to disease modeling

AbstractNeurological diseases are recognized as one of the most significant burdens of the modern society. Therefore, a new therapeutic approach applicable to nervous system represents priority of today’s medicine. A rapid development of stem cell technology in the last two decades introduced a possibility to regenerate disease-affected nervous tissue. In this vein, stem cells are envisioned as a replacement for lost neurons, a source of trophic support, a therapeutic vehicle, and as a tool for in vitro modeling. This article reviews the current concepts in stem cell-based therapy of neurological diseases and comments ongoing efforts aiming at clinical translation.

Review: Cellular repair strategies in Parkinson's disease

Parkinson's disease (PD) is the second most common neurodegenerative disorder after Alzheimer's disease, affecting 0.7% of the elderly population (defined as over 65 years of age). PD is clinically characterized by resting tremor, muscular rigidity, hypokinesia and postural instability. These motor symptoms result largely from the deficiency or dysfunction of dopaminergic neurons in the substantia nigra. Histopathological analysis reveals depletion of dopaminergic neurons as well as eosinophilic intracytoplasmic inclusions (Lewy bodies) in surviving neurons of the substantia nigra and other brain regions. The molecular pathogenesis is linked to protein misfolding by compromised alpha-synuclein and/or related proteins (synucleinopathy). Therefore, successful therapy of motor symptoms aims for the restoration of dopaminergic neurotransmission. Pharmacological drug treatment is usually effective only at an early stage of the disease but cannot halt progressive neuronal degeneration. With recent developments in stem cell technology, cell repair or replacement approaches came into focus. Here, we review new therapeutic strategies resulting from the innate propensity of the adult brain to generate new neurons, either by pharmacological stimulation of endogenous adult stem cell population or exogenous cell transplantation modalities.

Stamselnavorsing en -terapie: ’n oorsig oor die uitvoerbaarheid daarvan met meegaande etiese implikasies

Stem cell research and therapy: an overview of its feasibility with accompanying ethical implications Stem cell research is a breathtaking technology where man’s own cells are used to effect a cure for certain ailments. Stem cells are specially developed cells that contain the ability to develop into any cell in the body and to cure or replace defective cells. Researchers isolated stem cells from the human (and animal) embryo, foetus, umbilical cord, and adult body and developed them to stem cell lines for therapy. Astonishing results have already been shown. In some instances research- ers achieved full cure or at least huge improvement in animals (and in a few instances with human beings) with diabetes, Parkinson’s and Alzheimer’s diseases and auto-immune diseases. The broken spinal marrow of rats has also been repaired through this technique. The aim of this article is to give an overview of the technical feasibility and advancement of this field of research. It will become apparent from these facts that there are a large number of general and technical problems with the accompanying ethical implications, that will have to be resolved before this therapy can be applied. Problems that have been identified include the necessary destruction of the human embryo, the consideration of therapeutic cloning, the placing of human embryonic cells in embryos of primates such as apes and a series of technical problems in the development of stem cell technology.

Native Umbilical Cord Matrix Stem Cells Express Hepatic Markers and Differentiate Into Hepatocyte-like Cells

Umbilical cord matrix stem cells (UCMSCs) are able to differentiate into mesodermal and ectodermal lineages. The present study investigates the differentiation potential of human UCMSCs into hepatic lineage. We isolated human UCMSCs and characterized them in vitro by measuring their expansion potential, by assessing expression of mesenchymal stem cell (MSC) markers, and by evaluating their ability to differentiate into adipocytes and osteocytes. UCMSCs were thereafter subjected to a hepatogenic differentiation protocol. Expression of hepatic and MSC markers in differentiated cells was analyzed by reverse-transcription polymerase chain reaction, flow cytometry, and immunocytochemical assays and compared with undifferentiated UCMSCs and freshly isolated liver cells. UCMSCs were transplanted into livers of hepatectomized-SCID mice, and engraftment capacity was investigated by detection of human nucleus and mitochondria and human hepatic-specific proteins. In vitro expanded UCMSCs constitutively expressed markers of hepatic lineage, including albumin, alpha-fetoprotein, cytokeratin-19, connexin-32, and dipeptidyl peptidase IV. In vitro-differentiated UCMSCs exhibited hepatocyte-like morphology, up-regulated several hepatic markers, stored glycogen, produced urea, and exhibited an inducible CYP 3A4 activity. However, absence of some hepatic markers in differentiated UCMSCs, as HepPar1 or hepatocyte nuclear factor 4 (HNF-4), implied that their differentiation did not reach the level of mature hepatocytes. We also noticed that differentiated UCMSCs partially preserved MSC markers. Engraftment capacity of UCMSCs was observed, and expression of human albumin and alpha-fetoprotein was detected 2, 4, and 6 weeks after transplantation in mice livers, while cytokeratin 19 was completely down-regulated. We conclude that UCMSCs, with a newly demonstrated endodermic differentiation potential, might be an alternative source for liver-directed cell therapies.

Martin Evans: joint winner of 2007 Nobel Prize in medicine

Martin Evans: joint winner of 2007 Nobel Prize in medicine

Commercial Development of Stem Cell Technology: Lessons from the Past, Strategies for the Future

This paper presents historical and contemporary survey data on the commercial development of stem cell technology from the 1990s to the present day. We describe the first wave of industrial investment in hematopoietic stem cells during the 1990s and contrast this with the more recent expansion of the sector. In particular, we explore the cell types used, diseases targeted and business models adopted by firms. We conclude, by arguing that the commercial prospects for stem cell technologies remain highly uncertain and that innovative public policies should be adopted to prevent 'market failure'.

Genetic strategies for brain tumor therapy

Gene therapy is a potentially useful approach in the treatment of human brain tumors, which are notoriously refractory to conventional approaches. Most human clinical trials to date have been unsuccessful in terms of improving patient outcome. Recent improvements in viral vectors, the development of stem cell technology, and increased understanding of the mechanism of action of therapeutic transgenes provide hope that the next generation of gene therapeutics may show increased efficacy in treatment of this devastating disease.

Regeneration of the Rat Neonatal Intestine in Transplantation

Based on development of stem cell technology, newborn tissue, even undergoing cryopreservation, possesses promising potential as a donor source in the field of organ transplantation. However, the precise regeneration processes remains unclear. This study was designed to investigate the regenerative potential of newborn intestine with or without cryopreservation in the transplantation. Newborn rat intestines with or without cryopreservation were transplanted subcutaneously into the syngeneic host, and specimens were evaluated by histology, multiple immunostaining, and comprehensive gene expression analysis. We determined that newborn rat intestine possessed regenerative potential in the syngeneic host even after cryopreservation, where angiogenesis was induced early in the submucosa with subsequent maturation in the crypts. Furthermore, newborn intestinal graft could facilitate the survival of maturation-incompetent 10-day-old graft that lacked regenerating activity (P < 0.01, n = 13). Tissue aggregates from the maturation-incompetent graft underwent reconstitution of their histologic configuration in the presence of newborn intestinal aggregates. Comprehensive gene expression analysis showed that 37 genes were preferentially up-regulated, while 19 genes were down-regulated in the regenerating 10-day-old graft (supported by the newborn graft). Regeneration of newborn intestine is implicated in neo-angiogenesis in the host, and the newborn intestinal graft is capable of mediating the survival of the maturation-incompetent 10-day-old graft. Notwithstanding ethical and legal limitations in the clinic, these results may provide new insights into the regenerative role of newborn grafts.