ABSTRACT
The omentum is a sheet-like tissue attached to the greater curvature of the stomach and contains secondary lymphoid organs called milky spots. The omentum has been used for its healing potential for over 100 years by transposing the omental pedicle to injured organs (omental transposition), but the mechanism by which omentum helps the healing process of damaged tissues is not well understood. Omental transposition promotes expansion of pancreatic islets, hepatocytes, embryonic kidney, and neurons. Omental cells (OCs) can be activated by foreign bodies in vivo. Once activated, they become a rich source for growth factors and express pluripotent stem cell markers. Moreover, OCs become engrafted in injured tissues suggesting that they might function as stem cells.Omentum consists of a variety of phenotypically and functionally distinctive cells. To understand the mechanism of tissue repair support by the omentum in more detail, we analyzed the cell subsets derived from the omentum on immune and inflammatory responses. Our data demonstrate that the omentum contains at least two groups of cells that support tissue repair, immunomodulatory myeloid derived suppressor cells and omnipotent stem cells that are indistinguishable from mesenchymal stem cells. Based on these data, we propose that the omentum is a designated organ for tissue repair and healing in response to foreign invasion and tissue damage.
Subject(s)
Lung Injury/therapy , Omentum/physiology , Regeneration/physiology , Tissue Engineering/methods , Tissue Transplantation/methods , Totipotent Stem Cells/transplantation , Analysis of Variance , Animals , Bleomycin/toxicity , Blotting, Western , Bronchoalveolar Lavage , Cell Proliferation , DNA Primers/genetics , Flow Cytometry , Fluorescent Antibody Technique , Lung Injury/chemically induced , Mice , Mice, Inbred C57BL , Nitric Oxide Synthase Type II/metabolism , Omentum/cytology , Omentum/transplantation , Osteopontin/metabolism , Reverse Transcriptase Polymerase Chain Reaction , T-Lymphocytes/physiology , Tissue Transplantation/physiology , Totipotent Stem Cells/physiologyABSTRACT
Numerous studies have confirmed that stem cell therapy has significant potential for the regeneration of congenital and acquired heart diseases. The utilization of embryonic stem cells and induced pluripotent stem cells promises a possible generation and regeneration of all cardiovascular structures. On the one hand fetal and adult stem cells, e.g. endothelial progenitors, mesenchymal, hematopoietic, cardiac stem cells and myoblasts, possess limited potential for multilinear differentiation. On the other hand these cells have high paracrin activity and support with well-confirmed safety the reconstruction and formation of cardiovascular structures. On the visionary track towards an autonomously functioning autologous heart generated by tissue engineering, vascular, valvular and myocardial tissues have already been successfully created. This manuscript describes the possible stem cell sources for cardiovascular tissue engineering and evaluates their potency and safety from a medical and ethical point of view employing the data from systematic reviews (Medline database) and own investigations.
Subject(s)
Cardiovascular Diseases/surgery , Tissue Engineering/methods , Animals , Cell Differentiation/physiology , Embryonic Stem Cells/cytology , Embryonic Stem Cells/transplantation , Ethics, Medical , Ethics, Research , Guided Tissue Regeneration/ethics , Guided Tissue Regeneration/methods , Humans , Multipotent Stem Cells/cytology , Multipotent Stem Cells/transplantation , Pluripotent Stem Cells/cytology , Pluripotent Stem Cells/transplantation , Stem Cell Transplantation/methods , Tissue Engineering/ethics , Totipotent Stem Cells/cytology , Totipotent Stem Cells/transplantationABSTRACT
Stem cells are self-renewing cells capable of differentiating into multiple cell lines and are classified according to their origin and their ability to differentiate. Enormous potential exists in use of stem cells for regenerative medicine. To produce effective stem cell-based treatments for a range of diseases, an improved understanding of stem cell biology and better control over stem cell fate are necessary. In addition, the barriers to clinical translation, such as potential oncologic properties of stem cells, need to be addressed. With renewed government support and continued refinement of current stem cell methodologies, the future of stem cell research is exciting and promises to provide novel reconstructive options for patients and surgeons limited by traditional paradigms.
Subject(s)
Nuclear Transfer Techniques , Regenerative Medicine/methods , Regenerative Medicine/trends , Stem Cell Research , Stem Cell Transplantation , Stem Cells , Animals , Clinical Trials as Topic , Embryonic Stem Cells/transplantation , Humans , Multipotent Stem Cells/transplantation , Pluripotent Stem Cells/transplantation , Totipotent Stem Cells/transplantationABSTRACT
Planarians are well known for their remarkable regenerative capacity. This capacity to regenerate is thought to be due to the presence of totipotent somatic stem cells known as 'neoblasts', which have particular morphological characteristics. The totipotency of neoblasts was supported by Baguñà's experiment, which involved the introduction of donor cells into irradiated hosts. However, since Baguñà's experiment did not include the use of a phenotypic marker, the donor cells could not be traced. In the current study, a genetic mutant planarian, menashi, an eye-defective mutant that lacks the pigmented area in the eyes, was established. This planarian is excellent for tracing the fate of cells after their introduction into irradiated hosts. To investigate the differentiation potency more directly, a neoblast-rich fraction obtained from normal worms was transplanted into an X-ray-irradiated menashi strain. Planarians that survive X-ray irradiation were developed, and we observed the pigment of the area in the eyes of the regenerating planarians. This result suggests that the neoblast-rich fraction contains cells that can proliferate and differentiate. These cells can replace the cells and structures lost by X-ray irradiation and ablation, and they can also differentiate into eye pigment cells.
Subject(s)
Eye , Planarians/physiology , Regeneration/physiology , Totipotent Stem Cells/physiology , Animals , Eye/cytology , Eye Abnormalities/genetics , Mutation/genetics , Planarians/cytology , Planarians/genetics , Planarians/radiation effects , Totipotent Stem Cells/transplantationABSTRACT
Human pluripotent stem cells (PSCs) have the unique properties of being able to proliferate indefinitely in their undifferentiated state and of being able to differentiate into any somatic cell type. These cells are thus posited to be extremely useful for furthering our understanding of both normal and abnormal human development, providing a human cell preparation that can be used to screen for new reagents or therapeutic agents, and generating large numbers of differentiated cells that can be used for transplantation purposes. PSCs in culture have a specific morphology and they express characteristic surface antigens and nuclear transcription factors; thus, PSC culture is very specific and requires a core skill set for successful propagation of these unique cells. Specialized PSC training courses have been extremely valuable in seeding the scientific community with researchers that possess this skill set.
Subject(s)
Research Personnel/education , Teaching , Totipotent Stem Cells/cytology , Adult , Cell Differentiation , Cell Proliferation , Cells, Cultured , Financing, Government , Humans , National Institutes of Health (U.S.) , Research Personnel/organization & administration , Stem Cell Transplantation , Totipotent Stem Cells/physiology , Totipotent Stem Cells/transplantation , United StatesABSTRACT
Most therapeutic uses of stem cells demand that large numbers of cells are maintained in a Good Manufacturing Practice (GMP) facility, and envisage the development of a master depository from which a working bank of cells can be retrieved and differentiated into an appropriate phenotype for use. Likewise for gene- and drug-discovery processes, it is assumed that stable and genetically identical cells will eventually become available in large numbers. Critical for both of these assumptions is that the stem cells are stable during periods of amplification and differentiation. This review discusses the physiological features that must be assessed to measure stem cell stability, and proposes that genomic, epigenomic and mitochondrial markers, as well as functional measures of utility, should be considered. Recent findings suggesting that the level of cell stability is not homogeneous throughout all stem cells are also discussed.
Subject(s)
Cell Differentiation , Stem Cell Transplantation , Stem Cells/cytology , Cell Culture Techniques , Embryo, Mammalian/cytology , Humans , Totipotent Stem Cells/cytology , Totipotent Stem Cells/transplantationABSTRACT
Multipotential bone marrow stromal cells (MSCs) from wild-type (Wt) or apolipoprotein E deficient (Apoe(-/-)) mice were implanted into the cerebral ventricles of Apoe(-/-) mice. MSCs from Wt mice continued expressing apoE up to 6 months after implantation and were associated with enhanced novel object recognition and increased microtubule-associated protein 2 (MAP2) immunoreactivity in the dentate gyrus. These data show that MSCs can be used to distinguish developmental from post-developmental effects of a gene knockout and support their therapeutic potential for neurodegenerative diseases.
Subject(s)
Apolipoproteins E/metabolism , Stromal Cells/transplantation , Totipotent Stem Cells/transplantation , Animals , Animals, Newborn , Apolipoproteins E/genetics , Bone Marrow Cells/cytology , Bone Marrow Cells/metabolism , Bone Marrow Transplantation/methods , Cerebral Ventricles/metabolism , Cerebral Ventricles/surgery , Dentate Gyrus/metabolism , Female , Immunohistochemistry , Male , Mice , Mice, Inbred C57BL , Mice, Knockout , Microtubule-Associated Proteins/metabolism , Stromal Cells/cytology , Stromal Cells/metabolism , Totipotent Stem Cells/cytology , Totipotent Stem Cells/metabolismABSTRACT
AIM: The methods of inserting osseointegrated implants in atrophic edentulous ridges need first bone tissue graft and subsequently the insertion of fixtures. These methods need a long period of time before a prosthesis can be made: for this reason the researchers experimented different techniques to reduce times of prosthetic loading. Our experimented method makes it possible to shorten the prosthetic period notably. METHODS: During surgery an implant is inserted in the symphysis area and then it is removed by trephine bur having a diameter greater than 1 mm compared to the receiving site and by using irrigation with 4 degrees C physiological solution. After 75 days, the implant is loaded by temporary prosthesis and subsequently a final restoration is applied after 4-6 months, depending on bone quality. RESULTS: Our surgical method, in spite of being conditioned by the anatomical conformation of the edentulous ridge and anatomical limits, proved to be predictable and with the same success percentages as other surgical techniques used for morphological reconstruction of atrophic edentulous ridges, shortening prosthetic loading times notably. CONCLUSIONS: Despite the small number of patients treated and the short control period do not give enough elements to consider this new method applicable in all types of atrophies, our results confirm the validity of this technique if well used. In the future further studies and experimentations on greater number of patients and for at least 5 year follow-up are needed.
Subject(s)
Alveolar Ridge Augmentation/methods , Blood Platelets/metabolism , Bone Transplantation/methods , Dental Implantation, Endosseous/methods , Dental Implants , Growth Substances/therapeutic use , Jaw, Edentulous, Partially/surgery , Osseointegration , Adult , Aged , Antibiotic Prophylaxis , Atrophy , Batroxobin/pharmacology , Bone Marrow Transplantation , Dental Abutments , Female , Gels , Growth Substances/administration & dosage , Growth Substances/metabolism , Humans , Jaw, Edentulous, Partially/rehabilitation , Male , Middle Aged , Physical Stimulation , Totipotent Stem Cells/transplantation , Transplantation, Autologous , Wound HealingSubject(s)
Cloning, Organism/methods , Inheritance Patterns/genetics , Mice/physiology , Neurons, Afferent/cytology , Ploidies , Animals , Embryo Transfer , Female , Green Fluorescent Proteins , Male , Mice/genetics , Neurons, Afferent/physiology , Nuclear Transfer Techniques , Receptors, Odorant/metabolism , Totipotent Stem Cells/transplantation , Transgenes/genetics , beta-GalactosidaseSubject(s)
Nuclear Transfer Techniques , Regeneration/genetics , Stem Cell Transplantation/legislation & jurisprudence , Clone Cells , Genetic Therapy/legislation & jurisprudence , Genetic Therapy/trends , Humans , Missouri , Neurodegenerative Diseases/therapy , Pluripotent Stem Cells/cytology , Pluripotent Stem Cells/transplantation , Research/legislation & jurisprudence , Stem Cell Transplantation/trends , Totipotent Stem Cells/cytology , Totipotent Stem Cells/transplantationABSTRACT
Cells derived from embryonic mouse STO cell lines differentiate into hepatocytes when transplanted into the livers of nonimmunosuppressed dipeptidylpeptidase IV (DPPIV)-negative F344 rats. Within 1 day after intrasplenic injection, donor cells moved rapidly into the liver and were found in intravascular and perivascular sites; by 1 month, they were intrasinusoidal and also integrated into hepatic plates with approximately 2% efficiency and formed conjoint bile canaliculi. Neither donor cell proliferation nor host inflammatory responses were observed during this time. Detection of intrahepatic mouse COX1 mitochondrial DNA and mouse albumin mRNA in recipient rats indicated survival and differentiation of donor cells for at least 3 months. Mouse COX1 targets were also detected intrahepatically 4-9 weeks after STO cell injection into nonimmunosuppressed wild-type rats. In contrast to STO-transplanted rats, mouse DNA or RNA was not detectable in untreated or mock-transplanted rats or in rats injected with donor cell DNA. In cultured STO donor cells, DPPIV and glucose-6-phosphatase activities were observed in small clusters; in contrast, mouse major histocompatibility complex class I H-2Kq, H-2Dq, and H-2Lq and class II I-Aq markers were undetectable in vitro before or after interferon gamma treatment. Together with H-2K allele typing, which confirmed the Swiss mouse origin of the donor cells, these observations indicate that mouse-derived STO cell lines can differentiate along hepatocytic lineage and engraft into rat liver across major histocompatibility barriers.
Subject(s)
Cell Differentiation/physiology , Cell Lineage/physiology , Graft Survival/physiology , Liver/physiology , Totipotent Stem Cells/transplantation , Transplantation, Heterologous , Animals , Cell Line , Female , Immunosuppression Therapy , Liver/cytology , Mice , Rats , Rats, Inbred F344 , Totipotent Stem Cells/physiologyABSTRACT
The objectives of this study were to develop an in vitro culture system to optimize germ cell proliferation and to measure the potential of the cultured germ cells to produce mature spermatozoa after transplantation into a recipient. Donor germ cells isolated from ROSA26 male mice were cultured with a STO feeder cell layer in Dulbecco's minimal essential medium (DMEM) supplemented with fetal bovine serum (FBS), stem cell factor, leukemia inhibitory factor, basic fibroblast growth factor, insulin-like growth factor 1, interleukin-11, L-glutamine, sodium pyruvate, 2-mercaptoethanol, murine oncostatin M, and platelet-derived growth factor. Donor germ cells formed colonies in the primary cultures after 8-21 days. These cultured colonies were maintained for 4 weeks or longer without subculture and proliferated for up to 8 passages over a period of 3 months. These colonies had alkaline phosphatase activity and incorporated 5-bromo-2'-deoxyuridine. These colonies were positive partially when screened with antibody for germ cell nuclear antigen and c-kit. Germ cells cultured with this supplemented medium showed enhanced colonization vs controls cultured with DMEM and FBS. Cultured germ cells from Rosa26 donors were transplanted into testes and were identified by X-gal staining and histological screening. The cells cultured in the supplemented medium colonized the tubules and initiated spermatogenesis in the recipient mice. This is an improved method for culturing germ cells and may be useful in gene therapy and the production of transgenic animals.
Subject(s)
Cell Culture Techniques/methods , Spermatozoa/cytology , Stem Cell Transplantation/methods , Totipotent Stem Cells/transplantation , Animals , Cell Division , Male , Mice , Mice, Inbred Strains , Mice, Transgenic , Oligospermia/therapy , Testis/cytology , Totipotent Stem Cells/cytologySubject(s)
Endoderm/transplantation , Heart Failure/therapy , Stem Cell Transplantation , Stomach/cytology , Viscera/cytology , Animals , Cell Differentiation , Cell Division , Coculture Techniques , Endoderm/cytology , Endothelium, Vascular/cytology , Heart , Humans , Myocardium/cytology , Myocytes, Cardiac/cytology , Myocytes, Cardiac/physiology , Myocytes, Cardiac/transplantation , Paracrine Communication/physiology , Pluripotent Stem Cells/transplantation , Regeneration/physiology , Stem Cell Transplantation/legislation & jurisprudence , Stem Cell Transplantation/trends , Totipotent Stem Cells/transplantationABSTRACT
BACKGROUND AND AIMS: Embryonic stem (ES) cells have a pluripotent ability to differentiate into a variety of cell lineages in vitro. We have recently identified the emergence of cellular clusters within differentiated ES cell cultures by staining with dithizone (DTZ). DTZ is a zinc-chelating agent known to selectively stain pancreatic beta cells because of their high zinc content. The aim of the present study was to investigate the characteristics of DTZ-stained cellular clusters originating from ES cells. METHODS: Embryoid bodies (EBs), formed by a 5-day hanging drop culture of ES cells, were allowed to form outgrowths in the culture. The outgrowths were incubated in DTZ solution (final concentration, 100 microg/ml ) for 15 minutes before being examined microscopically. The gene expression of endocrine pancreatic markers was also analyzed by reverse transcriptase-polymerase chain reaction. In addition, insulin production was examined immunohistochemically, and its secretion was examined using enzyme-linked immunosorbent assay. RESULTS: DTZ-stained cellular clusters appeared after approximately 16 days in the EB culture and became more apparent by day 23. They were found to be immunoreactive to insulin and expressed pancreatic-duodenal homeobox 1 (PDX1), proinsulin 1, proinsulin 2, glucagon, pancreatic polypeptide, glucose transporter-2 (GLUT2), and islet-specific glucose-6-phosphatase catalytic subunit-related protein (IGRP) mRNA. They were also able to secrete detectable amounts of insulin. CONCLUSIONS: ES cell-derived DTZ-positive cellular clusters possess characteristics of the endocrine pancreas, including insulin secretion. Further, DTZ staining is a useful method for the identification of differentiated pancreatic islets developed from EBs in vitro.
Subject(s)
Cell Separation/methods , Chelating Agents , Diabetes Mellitus/therapy , Dithizone , Glucose-6-Phosphatase , Islets of Langerhans/drug effects , Stem Cell Transplantation/methods , Totipotent Stem Cells/transplantation , Animals , Cell Culture Techniques , Cell Separation/trends , Cells, Cultured , Female , Glucose Transporter Type 2 , Immunohistochemistry , Insulin/metabolism , Insulin Secretion , Islets of Langerhans/cytology , Islets of Langerhans/metabolism , Mice , Mice, Inbred Strains , Monosaccharide Transport Proteins/genetics , Proinsulin/genetics , Proteins/genetics , Stem Cell Transplantation/trends , Totipotent Stem Cells/cytology , Totipotent Stem Cells/metabolism , Zinc/metabolismABSTRACT
Spermatogonia are the male germ line stem cells. Their life long expansion is needed for permanent production of male germ cells. Spermatogonia are the only cells of the germ line, which proliferate in adulthood and offer interesting applications as they are potentially totipotent and immortal cells. This review presents some of the recent breakthroughs, which have led to a better understanding of spermatogonial physiology and opened new fields of basic research and of clinical applications in veterinary and medical science.
Subject(s)
Spermatogonia/transplantation , Animals , Cell Culture Techniques , Cryopreservation , Humans , Male , Spermatogonia/cytology , Totipotent Stem Cells/cytology , Totipotent Stem Cells/transplantationABSTRACT
Previously we have successfully delayed the onset of vasculitis and death in MRL Lpr/Lpr mice that are considered to have an autoimmune disease similar to human lupus erythematosus. Likewise, with the use of megadose human umbilical cord blood mononuclear cells, we were able to delay the onset of symptoms and death in SOD1 mice that carry a transgene for amyotrophic lateral sclerosis, considered by some to be an autoimune disease. A similar approach was utilized with NOD/LtJ type 1 diabetic mice. By administering megadoses of human umbilical cord blood mononuclear cells we were able to ameliorate the disease and improved the life span. This occurred to a greater extent than with bone marrow obtained from congenic mice. No immunosuppression was utilized in this study. This study raises the possibility of utilizing human cord blood mononuclear cells in conjunction with pancreatic islet transplantation.