902. [Derivation of germ cells from mouse embryonic stem cells in culture].
Mouse embryonic stem cells derive from the inner cell mass of the blastocyst and give rise to the three primitive embryonic layers, which later will form all the different tissue types of an adult. Embryonic stem cells are thus defined as totipotent cells. In vitro, these cells can give rise to all the somatic cells. Different laboratories have now shown that cultured embryonic stem cells can also differentiate into germline cells. By using the transcription factor Oct-4 as a tool for the visualization of germ cells, it has been shown the derivation of oocytes from mouse embryonic stem cells. These works should contribute to various areas, including therapeutic cloning which associates nuclear transfer and selective production of a specific cell type.
903. [Human embryonic stem cells: practical problems and scientific potentialities].
Human embryonic stem cells (hESCs) are derived from pre-implantation embryos given to research with the informed consent of the parents. These cells cannot give rise to a human being: they are not totipotent. They have an unlimited self-renewal capacity and they can generate the three embryonic germ layers, their respective derivatives and the extra-embryonic tissues: they are pluripotent. These cells represent an outstanding material for understanding functional genomics of not yet characterized human genes. They will be an important tool for pharmaceutical and clinical research.
904. [Extending preimplantation genetic diagnosis to HLA typing: the Paris experience].
作者: J Steffann.;N Frydman.;P Burlet.;N Gigarel.;E Feyereisen.;V Kerbrat.;G Tachdjian.;A Munnich.;R Frydman.
来源: Gynecol Obstet Fertil. 2005年33卷10期824-7页
Preimplantation genetic diagnosis (PGD) consists in the genetic analysis of one or two cells. These cells (blastomeres) are sampled from embryos, obtained by in vitro fertilization, at the third day of development. Since 1998, the bioethical laws (1994) and their decrees restricted PGD practices in France, strictly to the avoidance of the birth of a child affected with a genetic defect. In parallel, works on blood cord transplantation, taken at the birth of a compatible HLA sibling, showed very encouraging results, particularly for the treatment of Fanconi anemia. In 2001, Verlinsky et al., have reported the first PGD for Fanconi anaemia combined with HLA typing, allowing the birth of a healthy child, HLA-identical with his affected sister. The "designer baby" concept was born. The French law, which allowed PGD under specific conditions, i.e. when the genetic defect has been characterized in one parent at least, recently extended PGD to HLA typing when embryos are at risk of a genetic disorder. Article L.2131-4-1 (August 2004) allows the practice of HLA typing for PGD embryos when an elder sibling is affected with a genetic disorder and need stem cell transplantation. The HLA-matched offspring resulting from PGD can give cord blood at birth to supply the necessary therapy. This double selection give rise to serious ethical problems, but technical difficulties and legal restrictions will probably limit the development of such a procedure.
905. [Testicular tissue cryoconservation for prepubertal boy: indications and feasibility].
Childhood cancers below 15 years old represent almost 2000 new cases each year; their evolution is often favourable since about 75% of children will recover from their cancer. However, the gonad, in particular the testis, is very sensitive to radiotherapy and alkylant agents. Cryopreservation of semen can be proposed to young adults before starting the treatment; the 14-17 years period can be considered as a critical phase, because some boys have not achieved puberty. Thereby semen collection failures are frequent and semen quality is sometimes poor. In prepubertal boys, testicular stem cell banking is the only way of preserving the future fertility. Transplantation of testicular tissue was tempted for the first time in 1994: recipient mice produced gametes derived from testicular stem cells of donor mice, and could produce transgenic offspring. However, in the mouse, births after transplantation of cryopreserved stem cells were obtained only recently, in 2003. A lot of questions remain uncompletely resolved in animal models and of course in humans: stem cells enrichment, freezing-thawing protocols, transplantation procedure and corresponding health risks, represented by the possible re-introduction of malignant cells. Xenotransplantation or in vitro maturation of spermatogonia after thawing the testicular tissue could represent an alternative permitting to obtain malignant cell-free spermatozoa. However, a complete spermatogenesis in vitro has not yet been obtained, in any species.
907. [Neurogenesis in the pathologies of the nervous system].
Neural stem cells (NSCs) are the self-renewing, multipotent cells that generate neurons, astrocytes, and oligodendrocytes in the nervous system. Contrary to the long-held dogma, neurogenesis occurs in discrete areas of the adult brain, the hippocampus and the subventricular zone, and NSCs reside in the adult central nervous system. Recent studies have shown that neurogenesis is increased in the diseased brains, after strokes and traumatic brain injuries, and that new neuronal cells are generated at the sites of injury, where they replace some of the degenerated nerve cells. Thus, the central nervous system has the capacity to regenerate after injury. The contribution and function of the increased neurogenesis in the pathologies of the nervous system remain to be understood. The increased hippocampal neurogenesis may play a role in neuroadaptation, such as in memory troubles and depression, associated with these pathologies. The increased neurogenesis at the sites of injury may represent an attempt by the central nervous system to regenerate itself after injury. Newly generated neuronal cells at the sites of injury originate from the subventricular zone. Hence, strategies that would promote neurogenesis in the subventricular zone may promote neuronal repair after injury of the nervous system. In this manuscript, we will review the studies on neurogenesis in the pathologies of the nervous system.
909. [Cell therapy prospects in Duchenne muscular dystrophy].
Skeletal muscle is made of multinucleated postmitotic fibers which are the agents of contraction. These fibers arise from mononuclear precursor cells which fuse after having migrated from the somites to the site of myogenesis. The cascade of events which result in muscle differentiation is well known nowadays (Sabourin & Rudnicki, 2000). Some precursor cells present in adults muscle are termed satellite cells (Mauro, 1961), because of their location between the plasma membrane and the extracellular matrix of muscle fibers. Since they have a role in muscle growth and regeneration, these cells were the first candidates for treating muscle dystrophies. Despite a large corpus of positive experimental data about transplantation of these cells, no hard clinical result has been obtained to date, However these investigations have led to fruitful thinking about heterogeneity of muscle cell precursors and the possible ways to use them for therapy.
910. [Myoblast transplantation for heart failure: where are we heading?].
Cell therapy in cardiology is already a reality, as evidenced by the number of ongoing clinical trials. These studies entail administration of either skeletal myoblasts in patients with severe ischemic left ventricular dysfunction or of bone marrow-derived cells in patients with acute myocardial infarction and in whom cell therapy is an adjunct to a percutaneous revascularization procedure. The techniques of preparation, expansion and storage of myoblasts are now quite effective. The problem is simpler for bone marrow cells as in most studies, the procedure is limited to an iliac crest biopsy followed by reinjection of the crude, unfractionated bone marrow, as routinely done in clinical haematology since many years. The results of these studies are not yet fully available. Some of them have been enthusiastically reported to be positive but should be interpreted cautiously because of the usually small sample sizes and the common lack of randomisation and double-blind assessment of outcomes. Thus, the fact that cell therapy has now become a reality should not lead to underscore the yet unsettled fundamental issue, i.e., the ability of this novel mode of therapy to truly regenerate areas of necrotic myocardium and restore function in once akinetic territories. From this standpoint, cell therapy is still a dream. Since the beginning, it has been clear that myoblasts were exclusively committed to differentiate into myotubes, without any evidence for a phenotypic conversion into cardiomyocytes. Although the debate is more controversial for bone marrow cells, the reliance on accurate genetic methods of cell tracking has led to increasingly challenge the purported plasticity of these cells. This by no means implies that cell therapy does not exert beneficial effects that could be mediated by alternate mechanisms like limitation of remodelling of paracrine effects. The basic point is that neither skeletal myoblasts nor bone marrow cells fulfill the major criteria required for a true cardiac regeneration: a coupling of the grafted cells with those of the recipient myocardium and the subsequent generation of a contractile force. It is therefore critical to go on exploring other paths, among which embryonic stem cells are particularly attractive.
911. [Cardiac cellular therapy: from cells to the first clinical uses].
作者: J Roncalli.;B Leobon.;P Massabuau.;M Galinier.;A Parini.;A Pathak.;P Bourin.;A A Hagege.;P Menasche.;G Fournial.;J M Fauvel.
来源: Arch Mal Coeur Vaiss. 2005年98卷6期637-48页
Despite the improvement in revascularisation techniques, coronary artery disease remains the principal aetiology of cardiac failure in developed countries. The therapeutic management of cardiac failure has been improved over recent years, yet cardiac failure is still associated with significant morbidity and mortality. As cardiac transplantation lacks donors, techniques that allow myocardial regeneration represent an attractive alternative. To date, several types of cells are under study and are suitable for implantation into infarcted myocardium (myoblasts, medullary stem cells...). Following good preclinical study results, the first human cell therapy trials, using the intramyocardial route, have begun, in the course of aorto-coronary bypass surgery in patients with chronic ischaemic cardiopathy and little altered left ventricular function, and then in those with ventricular dysfunction. Different modes of administration of these cell therapy products are under study and could be envisaged in clinical situations such as just after infarction in order to improve ventricular remodelling with an intracoronary injection technique. As for every new treatment, there are numerous problems to resolve, from understanding the relevant mechanisms of cellular transplantation, to the secondary effects that it could entail. Nevertheless, cardiac cellular transplantation is expanding rapidly and with the evolution of techniques it allows a glimpse of a new field of treatment for cardiac failure.
913. [Systemic mastocytosis].
作者: O Fain.;J Stirnemann.;V Eclache.;S Barete.;P Casassus.;O Hermine.;O Lorholary.
来源: Presse Med. 2005年34卷9期681-7页
Systemic mastocytosis is characterized by abnormal mast cell proliferation in different organs. The 2001 consensus classification distinguishes in separate categories indolent systemic mastocytosis, systemic mastocytosis with concomitant blood disease, aggressive systemic mastocytosis and mast cell leukemia. Clinical manifestations are caused by tissue infiltration by proliferating mastocytes and by release of mediators. The principal organs affected are the skin, bones, digestive tract, liver, spleen and lymph nodes. Diagnosis of mastocytosis is based on appropriate stains (Giemsa, toluidine blue) and immunophenotype features (tryptase, CD117, also known as c-KIT and stem cell factor receptor). Serum tryptase levels reflect the total mast cell burden. Treatment must prevent release of mast cell mediators (histamine antagonists, cromolyn sodium, corticosteroids, or leukotriene-receptor inhibitors), limit bone involvement (bisphosphonates) and reduce the number of circulating mast cells (interferon, cladribine, or tyrosine kinase inhibitors). Enhanced understanding of the pathogenic mechanisms (mutation of c-kit and platelet-derived growth factor receptor alpha has led to the development of targeted treatments, including new inhibitors of tyrosine kinase and of nuclear factor Kappa B.
915. [Which stem cells for adult liver?].
While hepatocytes can be considered conceptually as unipotent stem cells, the presence of true stem or progenitor cells within adult livers has been largely debated. It is now accepted that the atypical ductular reaction observed in livers with sub-massive hepatitis represents the proliferation of hepatic progenitor cells similar to rat oval cells and able to differentiate towards the biliary and the hepatocytic lineage through intermediate progeny. In the normal liver, the identification of progenitor cells with a panel of markers including c-kit, CD34, Ov6, CK7, CK19, chromogranine A, CD56 remains difficult because these cells are very few and most of the markers are not specific. These progenitor cells could be located either within the canals of Hering or in periductular situation or both. Mechanisms leading to the activation and the proliferation of hepatic progenitor cells are still largely unknown: they involve growth factors as the stem cell factor, ligand of c-kit, cytokines, chemokines as SDF1 a and vagal or sympathetic innervatioñ. Other potential stem cells for liver could be hematopoietic stem cells from bone marrow. First publications have showed that hematopoietic stem cells were able to differentiate into hepatocytes and cholangiocytes and to yield high level engraftment of injured livers. However it appears now that this phenomenon is minimal or even absent in physiological and usual pathological conditions. It does occur in extreme experimental conditions either by true transdifferentiation or cell fusion. The shared property of stem cells and tumor cells to proliferate endlessly, rises the question of the potential role of progenitor cells in liver carcinogenesis. In a number of animal models of hepatocarcinogenesis, tumors originate from oval cells. The identification of progenitor cells close to murine oval cells in the human liver raises the hypothesis of a potential role of these cells in the development of human liver tumors. Liver progenitor cells have been identified morphologically and phenotypically in dysplastic foci of cirrhotic livers and hepatocellular adenomas. More generally speaking, typical hepatocellular carcinomas and cholangiocarcinomas are at the two ends of a spectrum which includes transitional-type tumors intermediate between hepatocellular carcinoma and cholangiocarcinoma and combined hepato-cellular cholangiocarcinoma; these intermediate and combined types can be more easily explained as deriving from progenitor cells. Despite the difficulties, the doubts and the potential dangers, new experimental modalities to obtain efficient repopulation of the liver from bone marrow stem cells are currently under study: exogenous administration of cytokines and chemokines involved in cell homing and differentiation or development of selective pressure strategies. Other cell types as intra-hepatic progenitor cells, bone marrow multipotent adult progenitor cells (MAPCs) or fetal hepatocytes could be alternative sources for liver cell therapy. Thus, progressing knowledge about stem cells in adult liver would allow to better understand mechanisms of hepatic homeostasia and regeneration and would open the way to cell-based therapy for liver diseases.
916. [Where have the neuronal stem cells of the subependymal zone gone in human beings?].
Stem cells are characterized by their ability for self-renewal (allowing them to be present throughout the entire life of the organism) and their ability to give rise to differentiated cells belong to one or more lineages. The strict definition of these cells is however still a matter of debate. There is new experimental evidence (including in human beings) that stem cells are present within the brain and may give rise to neurons. Ependymal cells have been proposed to play such a role. In fact, subependymal cells expressing GFAP would be more likely candidates. Such cells are observed in the brain of human beings. They are able to differentiate into neurons in vitro but such potential appears to be repressed in vivo.
917. [Cardiopulmonary function before and after cyclophosphamide treatment in severe systemic sclerosis: comparison of monthly intravenous bolus and autologous haematopoietic stem cell transplantation].
作者: C Toledano.;C Henegar.;D Ilie.;D Launay.;K Tiev.;I Marie.;Z Marjanovic.;J Emmerich.;J Cabane.;S Ménasché.;D Farge.
来源: Rev Med Interne. 2005年26卷6期444-52页
Cyclophosphamide in monthly intravenous bolus is used to treat severe forms of systemic sclerosis with pulmonary involvement. Since 1996, cyclophosphamide therapeutic intensification with autologous haematopoietic stem cells transplantation allowed significant improvement in skin and functional scores in severe systemic sclerosis. Cyclophosphamide potential cardiotoxicity in this setting has been questioned.
919. [Hope and limits in cell therapies].
The hematopoietic system is one of the best characterized human cellular system in which a multipotent adult stem cell is at the origin of all the cells of a tissue. These last years, the use of stem cells has given rise to many hopes in regenerative medicine, especially in diseases without efficient therapies. However the hematopoietic system in which stem cell transplantation has entered in clinical practice for many years has also shown the limits of these approaches. Especially the in vitro manipulation of hematopoietic stem cells remains a challenge which requires more fundamental knowledge on the biology of stem cells including self renewal and homing. Knowledge for other tissue system is even more preliminary but the same experimental strategy used for hematopoietic stem cell can be translated and may accelerate their use in cell therapies. Characterization of human adult pluripotent stem cells and the generation of human ES cells capable to differentiate towards several tissues have led to new hopes but the road to their use in therapies may be long and will require a lot of investment in basic biology.
920. [BK virus infection in a child after an hematopoietic stem cell transplantation].
作者: A Benketira.;R Tichit.;J Tenenbaum.;G Margueritte.;F Bernard.
来源: Arch Pediatr. 2005年12 Suppl 1卷S64-6页
Polyomavirus hominis 1, better known as BK virus (BKV), infects up to 90% of the general population. Significant clinical manifestations can be seen in immunocompromised patients. We report a case of haemorragic cystitis likely due to BKV in a child after allotransplantation of hematopoietic stem cells. A 10-year old boy with poor-prognosis acute T lymphoblastic leukaemia underwent cord blood allogeneic stem cell transplantation while in his first relapse. Macroscopic haematuria and low back pain occurred by day 95, in the context of acute graft versus host disease and pulmonary aspergillosis. Histopathologic examination showed a cytopathogenetic effect consistent with the diagnosis of BKV infection. Urinary PCR was positive for BKV. Treatment with cidofovir was followed by a marked improvement of urinary symptoms. The current understanding, diagnosis, and treatment of BKV-associated infection is discussed.
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