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1.
Cytotherapy ; 15(3): 255-62, 2013 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-23579058

RESUMO

BACKGROUND AIMS: Wide acceptance of the colony-forming unit (CFU) assay as a reliable potency test for stem cell products is hindered by poor inter-laboratory reproducibility. The goal of this study was to ascertain current laboratory practices for performing the CFU assay with an eye towards identifying practices that could be standardized to improve overall reproducibility. METHODS: A survey to evaluate current laboratory practices for performing CFU assays was designed and internationally distributed. RESULTS: There were 105 respondents to the survey, of whom 68% performed CFU assays. Most survey recipients specified that an automated rather than a manual cell count was performed on pre-diluted aliquots of stem cell products. Viability testing methods employed various stains, and when multiple sites used the same viability stain, the methods differed. Cell phenotype used to prepare working cell suspensions for inoculating the CFU assay differed among sites. Most respondents scored CFU assays at 14-16 days of incubation, but culture plates were read with various microscopes. Of 57 respondents, 42% had not performed a validation study or established assay linearity. Only 63% of laboratories had criteria for determining if a plate was overgrown with colonies. CONCLUSIONS: Survey results revealed inconsistent inter-laboratory practices for performing the CFU assay. The relatively low number of centers with validated CFU assays raises concerns about assay accuracy and emphasizes a need to establish central standards. The survey results shed light on numerous steps of the methodology that could be targeted for standardization across laboratories.


Assuntos
Terapia Baseada em Transplante de Células e Tecidos , Ensaio de Unidades Formadoras de Colônias/normas , Células-Tronco Hematopoéticas , Células-Tronco/citologia , Antígenos CD34/metabolismo , Transplante de Células-Tronco Hematopoéticas , Humanos , Padrões de Referência , Reprodutibilidade dos Testes
2.
J Biomed Biotechnol ; 2011: 172965, 2011.
Artigo em Inglês | MEDLINE | ID: mdl-22187520

RESUMO

Most anti-cancer immunotherapeutic strategies involving dendritic cells (DC) as vaccines rely upon the adoptive transfer of DC loaded with exogenous tumour-peptides. This study utilized human acute myeloid leukemia (AML) cells as progenitors from which functional dendritic-like antigen presenting cells (DLC) were generated, that constitutively express tumour antigens for recognition by CD8(+) T cells. DLC were generated from AML cell lines KG-1 and MUTZ-3 using rapid culture techniques and appropriate cytokines. DLC were evaluated for their cell-surface phenotype, antigen uptake and ability to stimulate allogeneic responder cell proliferation, and production of IFN-γ; compared with DC derived from normal human PBMC donors. KG-1 and MUTZ-3 DLC increased expression of CD80, CD83, CD86, and HLA-DR, and MUTZ-3 DLC downregulated CD14 and expressed CD1a. Importantly, both KG-1 and MUTZ-3-derived DLC promoted proliferation of allogeneic responder cells more efficiently than unmodified cells; neither cells incorporated FITC-labeled dextran, but both stimulated IFN-γ production from responding allogeneic CD8(+) T cells. Control DC produced from PBMC using the FastDC culture also expressed high levels of critical cell surface ligands and demonstrated good APC function. This paper indicates that functional DLC can be cultured from the AML cell lines KG-1 and MUTZ-3, and FastDC culture generates functional KG-1 DLC.


Assuntos
Técnicas de Cultura de Células/métodos , Células Dendríticas/metabolismo , Leucemia Mieloide Aguda/imunologia , Leucemia Mieloide Aguda/patologia , Análise de Variância , Antígenos CD/metabolismo , Linfócitos T CD8-Positivos/imunologia , Linfócitos T CD8-Positivos/metabolismo , Linhagem Celular Tumoral , Citocinas/metabolismo , Células Dendríticas/imunologia , Dextranos/química , Citometria de Fluxo , Fluoresceína-5-Isotiocianato/análogos & derivados , Fluoresceína-5-Isotiocianato/química , Antígenos de Histocompatibilidade Classe I/metabolismo , Humanos , Interferon gama/metabolismo , Leucócitos Mononucleares/imunologia , Leucócitos Mononucleares/patologia , Fenótipo
4.
Stem Cells ; 29(9): 1322-6, 2011 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-21739527

RESUMO

Hematopoietic stem cells (HSCs) have remained at the forefront of stem cell research for the past 50 years, since the therapeutic potential of bone marrow transplantation was realized. Uniquely, among stem and progenitor cells, research progress has been made in parallel between the laboratory benchtop and hospital bedside during this period. Integral to this work has been the role of the transfusion medicine services in the collection, storage, and processing of HSCs. The next decade promises to bring further developments: with new fields of cellular therapies, stem cell vaccination, and stem cell drug testing opening up. This article summarizes exciting areas of research concerning the behavior and potential clinical applications of HSCs. For the purposes of clarity, we describe in turn the trafficking and transfer of HSCs; ex vivo expansion of HSC units from different sources; and finally, applications of specifically selected subsets of hematopoietic cells and their progeny.


Assuntos
Transfusão de Sangue/métodos , Transplante de Células-Tronco Hematopoéticas/métodos , Células-Tronco Hematopoéticas/fisiologia , Humanos
6.
Expert Rev Hematol ; 4(2): 213-20, 2011 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-21495930

RESUMO

Hemopoietic stem-cell transplant patients may require intensive blood component support. Complications of transfusions include transmission of viral and bacterial infections, transfusion-associated graft-versus-host disease and transfusion-related acute lung injury. Alloimmunization to red cell antigens may cause difficulties in selecting compatible blood, while alloimmunization to HLA expressed on platelets may cause subsequent platelet transfusion refractoriness. It is essential to define robust transfusion policies and procedures and these should be regularly audited. This article reviews blood component transfusion in the setting of hemopoietic stem-cell transplant and specifically discusses the management of ABO-mismatched transplants, the prevention of cytomegalovirus transmission, the prevention of transfusion-associated graft-versus-host disease and the use of granulocyte transfusions.


Assuntos
Transfusão de Componentes Sanguíneos , Transplante de Células-Tronco Hematopoéticas , Incompatibilidade de Grupos Sanguíneos/imunologia , Doença Enxerto-Hospedeiro/etiologia , Doença Enxerto-Hospedeiro/prevenção & controle , Granulócitos/transplante , Antígenos HLA/imunologia , Humanos
8.
Transfusion ; 51(6): 1284-90, 2011 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-21133927

RESUMO

BACKGROUND: Most hematopoietic progenitor cell (HPC) products are infused or processed shortly after collection, but in some cases this may be delayed for up to 48 hours. A number of variables such as temperature and cell concentration are of critical importance for the integrity of HPCs during this time. STUDY DESIGN AND METHODS: We evaluated critical variables using cord blood HPC units that were divided equally and stored at 4 °C versus room temperature (RT) for up to 96 hours. Total nucleated cell (TNC) and mononuclear cell (MNC) counts, viable CD34+ cell counts, and CD45+ cell viability as well as colony-forming unit-granulocyte-macrophage (CFU-GM) present over time at each temperature were determined. RESULTS: Overall, the data indicate that with the exception of viable CD34+ cells, there was a significant decrease in each variable measured for 72 to 96 hours and, with the exception of viable CD34+ cells and CFU-GM, the reductions were significantly greater in RT units than 4 °C units. There was an increase in viable CD34+ count for units where TNC count was greater than 8.5 × 10(9) /L, compared with units where TNC count was less than 8.5 × 10(9) /L, that was different for each storage temperature. CONCLUSIONS: Cord blood HPC collections maintained at 4 °C retained higher TNC counts, MNC counts, and CD45+ cell viability over a 72- to 96-hour storage period.


Assuntos
Preservação de Sangue/métodos , Células Progenitoras de Granulócitos e Macrófagos/citologia , Células-Tronco Hematopoéticas/citologia , Antígenos CD34/metabolismo , Sangue Fetal/citologia , Temperatura
9.
Br J Haematol ; 147(1): 71-6, 2009 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-19681886

RESUMO

Haematopoietic stem cell transplantation (HSCT) is widely used to treat patients with a range of haematological and non-haematological disorders. Both bone marrow and peripheral blood stem cell collection are associated with morbidity and, very rarely, mortality. We investigated the information that exists to adequately inform donors about the relative merits of each procedure. We carried out a systematic review analysing data from six prospective randomised controlled trials of related donors and discuss here the merits and drawbacks of this approach. Registry data mostly describes patient outcome but stem cell donor registries collect and report information on unrelated donors which could easily be extended to related donors. Further well-designed, randomised studies are required.


Assuntos
Educação em Saúde/métodos , Transplante de Células-Tronco Hematopoéticas , Doadores de Tecidos/psicologia , Coleta de Tecidos e Órgãos/efeitos adversos , Seleção do Doador/normas , Medicina Baseada em Evidências/métodos , Humanos , Ensaios Clínicos Controlados Aleatórios como Assunto/métodos , Sistema de Registros , Coleta de Tecidos e Órgãos/métodos
10.
Cochrane Database Syst Rev ; (1): CD006406, 2009 Jan 21.
Artigo em Inglês | MEDLINE | ID: mdl-19160282

RESUMO

BACKGROUND: Haemopoietic stem cells can be collected from a donor either as a bone marrow harvest or by peripheral blood collection. Both techniques have risks for the donor. OBJECTIVES: The aim of this review was to identify the adverse effects of haemopoietic stem cell donation and to compare the tolerability and safety of the two methods. SEARCH STRATEGY: We searched bibliographic databases including the Cochrane Central Register of Controlled trials (CENTRAL) (The Cochrane Library 2008, issue 2), MEDLINE and EMBASE up to May 2008. We also searched reference lists of articles and contacted experts in the field. SELECTION CRITERIA: Randomised controlled trials enrolling haemopoietic stem cell donors and evaluating the different methods of donating haemopoietic stem cells were eligible. DATA COLLECTION AND ANALYSIS: Two authors independently screened studies for inclusion. We extracted data and evaluated methodological quality. Quantitative analysis was not possible for most outcomes, but where used we preferred random-effects models due to the variability between the included studies. MAIN RESULTS: Six trials (807 donors) were eligible: all were substudies, or constituent parts of, larger randomised controlled trials of bone marrow and peripheral blood stem cell allogeneic transplantation. No included trial was designed solely to measure and assess the experience of stem cell donors. The donors in all studies were related to the stem cell recipient. Overall, both types of donors experienced pain subsequent to donation, and psychological morbidity. The trend was for bone marrow donors to experience more pain at the donation site, more overall adverse events, and more days of restricted activity. They were also more likely to require hospitalisation than peripheral blood stem cell donors. In contrast, peripheral blood stem cell donors experienced more pain prior to donation, which may be related to the pre-donation administration of granulocyte colony stimulating factor (G-CSF). The methodological quality of the studies was poor and indicated limitations due to the risk of selection and attrition bias. The proportion of donors from the parent trial not included in the donor substudies was also inadequately explained. AUTHORS' CONCLUSIONS: The different short-term morbidities associated with each type of haemopoietic stem cell donation were clear, with bone marrow donors experiencing more pain and more restriction post-donation than peripheral blood donors. However, the studies were limited by their methodological quality, failure to provide long-term follow up (for which larger numbers of donors would be required) and a failure to apply consistent measures of quality of life in a way which allows more meaningful evaluation across studies.


Assuntos
Células da Medula Óssea , Dor Pós-Operatória/etiologia , Doadores de Tecidos , Coleta de Tecidos e Órgãos/efeitos adversos , Ansiedade/etiologia , Fator Estimulador de Colônias de Granulócitos/administração & dosagem , Mobilização de Células-Tronco Hematopoéticas/efeitos adversos , Células-Tronco Hematopoéticas , Humanos , Ensaios Clínicos Controlados Aleatórios como Assunto , Doadores de Tecidos/psicologia , Coleta de Tecidos e Órgãos/métodos , Coleta de Tecidos e Órgãos/mortalidade
11.
Hematol Oncol Stem Cell Ther ; 2(2): 311-9, 2009.
Artigo em Inglês | MEDLINE | ID: mdl-20118054

RESUMO

JACIE was initiated as a small pilot project in Spain in 2000 and launched as a formal Europe-wide inspection program in January 2004. Since 2000, over 150 applications for accreditation have been received by the JACIE Office and more than 130 inspections have been completed in European centers and facilities. Almost all of these were found to be functioning at a high level of excellence, with the majority having only minor deficiencies in compliance with the standards. In one-third of centers there were more significant deficiencies. The most common deficiencies were in quality management. Following correction of deficiencies 86 centers have to date achieved full accreditation and many more are nearing the completion of the process. Implementation of JACIE involves a significant investment of time and resources by applicant centers. The majority require at least 18 months to prepare for accreditation and 85% have needed to employ a quality manager and/or data manager on an ongoing basis. However, all centers felt their program had benefited from the implementation of JACIE. JACIE is also working closely with other international organisations related to cellular therapy as part of the Alliance for the Harmonisation of Cell Therapy Accreditation (AHCTA), which is examining the differences in existing standards and aiming to develop international standards for all aspects of stem cell transplantation. In particular the requirements for safety of imported tissues and cells has emphasised the need for global harmonisation. The recent implementation of Directive 2004/23/EC and the associated Commission Directives 2006/17/EC and 2006/86/EC has provided an impetus for the implementation of JACIE in European Union (EU) member states. It will be important in the future to examine how JACIE can co-operate with the EU Competent Authorities (CA) to ease the burden of the inspection process for haemopoietic stem cell (HSC) transplant programs.


Assuntos
Acreditação/normas , Terapia Baseada em Transplante de Células e Tecidos/normas , Neoplasias/terapia , Qualidade da Assistência à Saúde/normas , Transplante de Células-Tronco , União Europeia , Humanos
13.
Transfusion ; 48(7): 1495-501, 2008 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-18373639

RESUMO

Granulocyte-colony-stimulating factor (G-CSF) is used for the mobilization of hemopoietic stem cells in healthy donors. It has a number of common side effects such as bone pain, which resolve rapidly after administration is discontinued. Recent publications have raised concern that it might act as a trigger for the development of hematologic malignancy in susceptible individuals, possibly by causing genomic instability, but to date there is no evidence that healthy volunteer donors who receive G-CSF are at any increased risk. Ongoing studies aim to confirm whether or not G-CSF can cause chromosomal abnormalities in healthy donors. In the UK, the British Bone Marrow Registry and Anthony Nolan Trust give G-CSF to donors who have agreed to donate peripheral blood stem cells. It is recommended by the UK Registries at present that all stem cell donors are given updated information explaining the current uncertainties with regard to the use of G-CSF before they give informed consent to its administration. This information is based on a statement agreed by the World Marrow Donor Association for use by individual donor registries. Further, it is our current practice that all donors who have received G-CSF, as well as marrow donors who do not, should be under regular review for at least 10 years to allow the occurrence of any long-term adverse events to be documented.


Assuntos
Doadores de Sangue , Fator Estimulador de Colônias de Granulócitos/efeitos adversos , Mobilização de Células-Tronco Hematopoéticas , Osso e Ossos/efeitos dos fármacos , Osso e Ossos/fisiopatologia , Fator Estimulador de Colônias de Granulócitos/administração & dosagem , Cefaleia/induzido quimicamente , Transplante de Células-Tronco Hematopoéticas , Humanos , Náusea/induzido quimicamente , Dor/induzido quimicamente , Sistema de Registros/estatística & dados numéricos , Reino Unido , Vômito/induzido quimicamente
14.
Transfusion ; 48(3): 546-9, 2008 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-18067495

RESUMO

BACKGROUND: Clinical transplant outcome with umbilical cord blood (UCB) as source of hematopoietic progenitor cells (HPCs) is, among other factors, determined by the total number of viable nucleated cells and/or CD34+ cells in the unit. Quantitative and qualitative losses by processing and cryopreservation and by thawing and washing before transfusion may occur, however. Another reason for a discrepancy between the number of cells in the unit released by the cord blood bank and found in the transplant center may be technical differences in cell counting methods between the two sites. STUDY DESIGN AND METHODS: With the collaborative group for Biomedical Excellence for Safer Transfusion (BEST), an interlaboratory exercise was conducted among nine sites for thawed UCB variables: total nucleated cells, CD34+ cells, viability, and HPC cultures. Three frozen UCB samples were shipped, with instructions for thawing, counting, and HPC plating. RESULTS: Unexpectedly samples arrived at all nine receiving centers without detectable hematopoietic progenitor colony-forming cells. Nevertheless, wide interlaboratory ranges for viability were obtained. The proportion of viable cells was found higher with manual methods, but all viability assays used in the study overestimated functional progenitor cells. CONCLUSIONS: The results underscore the complexity of evaluation of frozen-thawed cord blood cells and the need for standardization of assessment.


Assuntos
Sangue Fetal/citologia , Células-Tronco Hematopoéticas/citologia , Antígenos CD34/sangue , Bancos de Sangue , Transfusão de Sangue/métodos , Transfusão de Sangue/normas , Contagem de Células , Sobrevivência Celular/fisiologia , Sangue Fetal/metabolismo , Hematologia/métodos , Células-Tronco Hematopoéticas/metabolismo , Humanos , Laboratórios/normas
16.
J Clin Apher ; 22(5): 258-64, 2007.
Artigo em Inglês | MEDLINE | ID: mdl-17823946

RESUMO

The publication of new standards for terminology and labeling marks an important step in ensuring consistency and traceability of cellular therapies at the global level. However, it is only with the widespread implementation of the standard that the benefits can be truly realized. This paper provides guidance on the practical aspects of adopting these new standards for organizations with differing current levels of computerization. It discusses project management, equipment, licensing, and validation topics.


Assuntos
Bancos de Sangue/normas , Guias de Prática Clínica como Assunto/normas , Rotulagem de Produtos/normas , Transplantes/normas , Transfusão de Sangue/normas , Fidelidade a Diretrizes , Humanos , Cooperação Internacional , Controle de Qualidade , Software
19.
Br J Haematol ; 137(5): 401-8, 2007 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-17488485

RESUMO

Robin Coombs was the last survivor of the distinguished group of immunologists that included Philip Gell, John Humphrey, John Marrack, Peter Medawar and Robert White and who were responsible for the renaissance of British Immunology after the Second World War. He is best remembered for describing the antiglobulin test that bears his name. The antiglobulin test revolutionised the diagnosis of haemolytic diseases and the compatibility testing of blood for transfusion. In all, Coombs authored over 200 scientific papers. Haemagglutination reactions became widely used in the diagnosis of a range of infectious agents. Together with Philip Gell, he devised the classification of allergic reactions; these were published in the textbook "Clinical Aspects of Immunology", which he and Gell first edited in 1963 and which became the leading textbook on medical immunology. Robin Coombs was also one of the founders of the British Society for Immunology.


Assuntos
Testes de Coagulação Sanguínea/história , Transfusão de Sangue/história , Alergia e Imunologia/história , História do Século XX , Humanos , Reino Unido
20.
Stem Cells ; 25(8): 2087-93, 2007 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-17510215

RESUMO

Umbilical cord blood (UCB) is an important source of hematopoietic stem cells for transplantation. Although UCB is often collected from unrelated donors, directed umbilical cord blood (DCB) from sibling donors also provides an important source of UCB for transplantation. This report summarizes the experience in collection, testing, storage, and transplantation of DCB units by the National Blood Service for England and North Wales over 10 years. Eligibility for collection was based on an existing sibling suffering from a disease that may be treated by stem cell transplantation or a family history that could result in the birth of a sibling with a disease that could be treated by stem cell transplantation. Collections were made on the provision that the sibling's clinician was willing to financially support the collection and to take responsibility for medical review of the mother and potential recipient. Given the high investment in UCB banking and the introduction of new regulations and mandatory licensing under the European Union Tissues and Cells Directive and those proposed in the U.S., this report details the procedures that we have used for DCB donations, the outcome data where donations have been used for transplantation, and it provides some timely recommendations for best practices. Disclosure of potential conflicts of interest is found at the end of this article.


Assuntos
Doadores de Sangue , Transplante de Células-Tronco de Sangue do Cordão Umbilical/métodos , Doação Dirigida de Tecido , Sangue Fetal/citologia , Irmãos , Armazenamento de Sangue/métodos , Criança , Pré-Escolar , Inglaterra , Feminino , Sangue Fetal/microbiologia , Seguimentos , Doenças Hematológicas/epidemiologia , Doenças Hematológicas/terapia , Humanos , Estudos Retrospectivos , Transplante/mortalidade , Transplante Homólogo , Resultado do Tratamento
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