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  1. Article ; Online: Preservation of critical quality attributes of mesenchymal stromal cells in 3D bioprinted structures by using natural hydrogel scaffolds.

    Martorell, Lluís / López-Fernández, Alba / García-Lizarribar, Andrea / Sabata, Roger / Gálvez-Martín, Patricia / Samitier, Josep / Vives, Joaquim

    Biotechnology and bioengineering

    2023  Volume 120, Issue 9, Page(s) 2717–2724

    Abstract: Three dimensional (3D) bioprinting is an emerging technology that enables complex spatial modeling of cell-based tissue engineering products, whose therapeutic potential in regenerative medicine is enormous. However, its success largely depends on the ... ...

    Abstract Three dimensional (3D) bioprinting is an emerging technology that enables complex spatial modeling of cell-based tissue engineering products, whose therapeutic potential in regenerative medicine is enormous. However, its success largely depends on the definition of a bioprintable zone, which is specific for each combination of cell-loaded hydrogels (or bioinks) and scaffolds, matching the mechanical and biological characteristics of the target tissue to be repaired. Therefore proper adjustment of the bioink formulation requires a compromise between: (i) the maintenance of cellular critical quality attributes (CQA) within a defined range of specifications to cell component, and (ii) the mechanical characteristics of the printed tissue to biofabricate. Herein, we investigated the advantages of using natural hydrogel-based bioinks to preserve the most relevant CQA in bone tissue regeneration applications, particularly focusing on cell viability and osteogenic potential of multipotent mesenchymal stromal cells (MSCs) displaying tripotency in vitro, and a phenotypic profile of 99.9% CD105
    MeSH term(s) Hydrogels/chemistry ; Osteogenesis ; Gelatin/chemistry ; Tissue Engineering/methods ; Mesenchymal Stem Cells ; Fibrin/metabolism ; Tissue Scaffolds/chemistry ; Bioprinting/methods ; Printing, Three-Dimensional
    Chemical Substances Hydrogels ; Gelatin (9000-70-8) ; Fibrin (9001-31-4)
    Language English
    Publishing date 2023-03-23
    Publishing country United States
    Document type Journal Article ; Research Support, Non-U.S. Gov't
    ZDB-ID 280318-5
    ISSN 1097-0290 ; 0006-3592
    ISSN (online) 1097-0290
    ISSN 0006-3592
    DOI 10.1002/bit.28381
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  2. Article ; Online: Variable readthrough responsiveness of nonsense mutations in hemophilia A.

    Martorell, Lluis / Cortina, Vicente / Parra, Rafael / Barquinero, Jordi / Vidal, Francisco

    Haematologica

    2020  Volume 105, Issue 2, Page(s) 508–518

    Abstract: Readthrough therapy relies on the use of small molecules that enable premature termination codons in mRNA open reading frames to be misinterpreted by the translation machinery, thus allowing the generation of full-length, potentially functional proteins ... ...

    Abstract Readthrough therapy relies on the use of small molecules that enable premature termination codons in mRNA open reading frames to be misinterpreted by the translation machinery, thus allowing the generation of full-length, potentially functional proteins from mRNA carrying nonsense mutations. In patients with hemophilia A, nonsense mutations potentially sensitive to readthrough agents represent approximately 16% of the point mutations. The aim of this study was to measure the readthrough effect of different compounds and to analyze the influence of premature termination codon context in selected nonsense mutations causing hemophilia A. To this end, primary fibroblasts from three patients with hemophilia A caused by nonsense mutations (p.W1586X, p.Q1636X and p.R1960X) and Chinese hamster ovary (CHO) cells transfected with 12 different plasmids encoding mutated
    MeSH term(s) Animals ; CHO Cells ; Codon, Nonsense ; Cricetinae ; Cricetulus ; Hemophilia A/drug therapy ; Hemophilia A/genetics ; Humans ; RNA, Messenger/genetics
    Chemical Substances Codon, Nonsense ; RNA, Messenger
    Language English
    Publishing date 2020-01-31
    Publishing country Italy
    Document type Journal Article ; Research Support, Non-U.S. Gov't
    ZDB-ID 2333-4
    ISSN 1592-8721 ; 0017-6567 ; 0390-6078
    ISSN (online) 1592-8721
    ISSN 0017-6567 ; 0390-6078
    DOI 10.3324/haematol.2018.212118
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  3. Article ; Online: Beyond chimerism analysis: methods for tracking a new generation of cell-based medicines.

    Vives, Joaquim / Casademont-Roca, Aina / Martorell, Lluís / Nogués, Núria

    Bone marrow transplantation

    2020  Volume 55, Issue 7, Page(s) 1229–1239

    Abstract: The analysis of chimerism is crucial to determine the status of patients receiving hematopoietic stem cell transplantation. The variety of relevant techniques available today range from those that analyse nucleic acids (i.e. polymerase chain reaction ... ...

    Abstract The analysis of chimerism is crucial to determine the status of patients receiving hematopoietic stem cell transplantation. The variety of relevant techniques available today range from those that analyse nucleic acids (i.e. polymerase chain reaction based, next generation sequencing) and cellular phenotype (i.e. flow cytometry) to sophisticated imaging (particularly multimodal imaging using labelling agents). However, current developments of advanced therapies bring chimerism studies into a new dimension in which methods for detection of donor cells in the patient need to adapt to a wider range of cell- and gene-based medicines, routes of administration, target organs and pathologies. Herein we describe and analyze the toolkit of suitable labelling and detection methodologies with actual examples along with a discussion on challenges ahead and potential solutions. Remarkably, existing methods commonly used in chimerism analysis are suitable for use with new cell- and gene-based medicines. Indeed, new developments may facilitate the evolution and combination of such methodologies to the use of non-invasive and highly informative approaches.
    MeSH term(s) Chimerism ; Flow Cytometry ; Hematopoietic Stem Cell Transplantation ; Humans ; Polymerase Chain Reaction ; Transplantation Chimera
    Language English
    Publishing date 2020-02-05
    Publishing country England
    Document type Journal Article ; Research Support, Non-U.S. Gov't ; Review
    ZDB-ID 632854-4
    ISSN 1476-5365 ; 0268-3369 ; 0951-3078
    ISSN (online) 1476-5365
    ISSN 0268-3369 ; 0951-3078
    DOI 10.1038/s41409-020-0822-8
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  4. Article ; Online: Use of Multipotent Mesenchymal Stromal Cells, Fibrin, and Scaffolds in the Production of Clinical Grade Bone Tissue Engineering Products.

    Vives, Joaquim / Rodríguez, Luciano / Coca, Maria Isabel / Reales, Laura / Cabrera-Pérez, Raquel / Martorell, Lluís

    Methods in molecular biology (Clifton, N.J.)

    2020  Volume 2286, Page(s) 251–261

    Abstract: Tissue engineering products (TEP) are a new type of medicines resulting from the combination of cells, scaffolds, and/or signalling factors, which can be used for the regeneration of damaged tissues thus opening new avenues for the treatment of complex ... ...

    Abstract Tissue engineering products (TEP) are a new type of medicines resulting from the combination of cells, scaffolds, and/or signalling factors, which can be used for the regeneration of damaged tissues thus opening new avenues for the treatment of complex conditions. However, such combination of biologically active elements, particularly living cells, poses an unprecedented challenge for their production under pharmaceutical standards.In the methods presented here, we formulated two types of TEP based on the use of multipotent mesenchymal stromal cells with osteogenic potential combined with osteoinductive and osteoconductive bony particles from tissue bank embedded in a fibrin hydrogel that, altogether, can induce the generation of new tissue while adapting to the diverse architecture of bony defects. In agreement with pharmaceutical quality and regulatory requirements, procedures presented herein can be performed in compliance with current good manufacturing practices and be readily implemented in straightforward facilities at hospitals and academic institutions.
    MeSH term(s) Bone Regeneration ; Cells, Cultured ; Fibrin Tissue Adhesive/chemistry ; Fibrin Tissue Adhesive/pharmacology ; Humans ; Hydrogels/chemistry ; Hydrogels/pharmacology ; Mesenchymal Stem Cells/cytology ; Mesenchymal Stem Cells/drug effects ; Mesenchymal Stem Cells/physiology ; Primary Cell Culture/methods ; Tissue Engineering/methods ; Tissue Scaffolds/chemistry
    Chemical Substances Fibrin Tissue Adhesive ; Hydrogels
    Language English
    Publishing date 2020-07-23
    Publishing country United States
    Document type Journal Article ; Research Support, Non-U.S. Gov't
    ISSN 1940-6029
    ISSN (online) 1940-6029
    DOI 10.1007/7651_2020_280
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  5. Article ; Online: Virus-Specific T Cells From Cryopreserved Blood During an Emergent Virus Outbreak for a Potential Off-the-Shelf Therapy.

    Mora-Buch, Rut / Tomás-Marín, Maria / Enrich, Emma / Antón-Iborra, Mireia / Martorell, Lluís / Valdivia, Elena / Lara-de-León, Ana Gabriela / Aran, Gemma / Piron, Maria / Querol, Sergi / Rudilla, Francesc

    Transplantation and cellular therapy

    2023  Volume 29, Issue 9, Page(s) 572.e1–572.e13

    Abstract: During the first outbreak of an emergent virus, methods need to be developed to rapidly establish suitable therapies for patients with high risk of severe disease caused by the pathogen. Considering the importance of the T-cell response in controlling ... ...

    Abstract During the first outbreak of an emergent virus, methods need to be developed to rapidly establish suitable therapies for patients with high risk of severe disease caused by the pathogen. Considering the importance of the T-cell response in controlling viral infections, adoptive cell therapy with virus-specific T cells has been used as a safe and effective antiviral prophylaxis and treatment for immunocompromised patients. The main objective of this study was to establish an effective and safe method to cryostore whole blood as starting material and to adapt a T-cell activation and expansion protocol to generate an off-the-shelf antiviral therapeutic option. Additionally, we studied how memory T-cell phenotype, clonality based on T-cell receptor, and antigen specificity could condition characteristics of the final expanded T-cell product. Twenty-nine healthy blood donors were selected from a database of convalescent plasma donors with a confirmed history of SARS-CoV-2 infection. Blood was processed using a fully automated, clinical-grade, and 2-step closed system. Eight cryopreserved bags were advanced to the second phase of the protocol to obtain purified mononucleated cells. We adapted the T-cell activation and expansion protocol, without specialized antigen-presenting cells or presenting molecular structures, in a G-Rex culture system with IL-2, IL-7, and IL-15 cytokine stimulation. The adapted protocol successfully activated and expanded virus-specific T cells to generate a T-cell therapeutic product. We observed no major impact of post-symptom onset time of donation on the initial memory T-cell phenotype or clonotypes resulting in minor differences in the final expanded T-cell product. We showed that antigen competition in the expansion of T-cell clones affected the T-cell clonality based on the T-cell receptor β repertoire. We demonstrated that good manufacturing practice of blood preprocessing and cryopreserving is a successful procedure to obtain an initial cell source able to activate and expand without a specialized antigen-presenting agent. Our 2-step blood processing allowed recruitment of the cell donors independently of the expansion cell protocol timing, facilitating donor, staff, and facility needs. Moreover, the resulting virus-specific T cells could be also banked for further use, notably maintaining viability and antigen specificity after cryopreservation.
    MeSH term(s) Humans ; COVID-19/epidemiology ; COVID-19/therapy ; COVID-19 Serotherapy ; SARS-CoV-2 ; T-Lymphocytes ; Cryopreservation ; Disease Outbreaks ; Antiviral Agents
    Chemical Substances Antiviral Agents
    Language English
    Publishing date 2023-06-06
    Publishing country United States
    Document type Journal Article ; Research Support, Non-U.S. Gov't
    ZDB-ID 3062231-1
    ISSN 2666-6367
    ISSN (online) 2666-6367
    DOI 10.1016/j.jtct.2023.06.001
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    Zs.A 5082: Show issues Location:
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  6. Article ; Online: Improving cord blood typing with next-generation sequencing: impact of allele-level HLA and NIMA determination on their selection for transplantation.

    Enrich, Emma / Vidal, Francisco / Corrales, Irene / Campos, Eva / Borràs, Nina / Martorell, Lluís / Sánchez, Mar / Querol, Sergi / Rudilla, Francesc

    Bone marrow transplantation

    2020  Volume 55, Issue 8, Page(s) 1623–1631

    Abstract: Allele-level HLA compatibility in cord blood transplantation, together with noninherited maternal antigen or NIMA matching, have been associated with better transplant outcomes. The aim of this work is to develop a cost-efficient high-resolution HLA ... ...

    Abstract Allele-level HLA compatibility in cord blood transplantation, together with noninherited maternal antigen or NIMA matching, have been associated with better transplant outcomes. The aim of this work is to develop a cost-efficient high-resolution HLA typing strategy based on next-generation sequencing to improve the quality of the Barcelona Cord Blood Bank's inventory, and to investigate the impact of high-resolution HLA typing and NIMA determination on the preferential selection of cord blood for transplantation. In this line, the developed strategy was validated and the HLA-A, -B, -C, -DRB1, and -DQB1 genes of 5000 cord blood units and 2500 of their associated maternal samples were typed. Subsequently, three study groups of 2012 units each were monitored for up to 2 years: (1) units with high-resolution and maternal HLA typing, (2) units with high-resolution but not maternal typing, and (3) units typed at low-resolution for class I and only high-resolution for HLA-DRB1. Despite a trend toward a greater selection of units with high-resolution typing, no significant impact of these variables was observed. These results highlight the need for evidence-based and globally accepted criteria for cord blood selection, together with the necessity to improve the accessibility of clinicians to donor registry's data.
    MeSH term(s) Alleles ; Blood Grouping and Crossmatching ; HLA-DRB1 Chains ; High-Throughput Nucleotide Sequencing ; Histocompatibility Testing ; Humans
    Chemical Substances HLA-DRB1 Chains
    Language English
    Publishing date 2020-04-17
    Publishing country England
    Document type Journal Article
    ZDB-ID 632854-4
    ISSN 1476-5365 ; 0268-3369 ; 0951-3078
    ISSN (online) 1476-5365
    ISSN 0268-3369 ; 0951-3078
    DOI 10.1038/s41409-020-0890-9
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  7. Article ; Online: HLA-A, -B, -C, -DRB1, and -DQB1 allele and haplotype frequencies: An analysis of umbilical cord blood units at the Barcelona Cord Blood Bank.

    Enrich, Emma / Campos, Eva / Martorell, Lluís / Herrero, María José / Vidal, Francisco / Querol, Sergi / Rudilla, Francesc

    HLA

    2019  Volume 94, Issue 4, Page(s) 347–359

    Abstract: Allele-level HLA compatibility in cord blood transplantation has been associated with better transplant outcomes and is recommended as a selection criterion. It is also a crucial aspect for other therapeutic applications involving cord blood-derived ... ...

    Abstract Allele-level HLA compatibility in cord blood transplantation has been associated with better transplant outcomes and is recommended as a selection criterion. It is also a crucial aspect for other therapeutic applications involving cord blood-derived cells. Determination of high-resolution HLA frequencies is an important step towards improving the quality of cord blood banks. We analyzed HLA-A, -B, -C, -DRB1, and -DQB1 allele frequencies in 5458 high-quality cord blood units from the Barcelona Cord Blood Bank and identified 275 class I and 121 class II HLA alleles. A*02:01, B*44:03, C*07:01, DRB1*07:01 and DQB1*03:01 were the most frequent alleles at each locus. We detected 26 novel alleles and were able to determine the presence or absence of some null alleles, including C*04:09N, in a large number of units. We also analyzed maternal HLA typing information for 1877 units to determine real haplotype frequencies and linkage disequilibrium. A*29:02-B*44:03-C*16:01-DRB1*07:01-DQB1*02:02 was the most frequent HLA haplotype and the DRB1-DQB1 gene pair contained the two-locus haplotypes with the strongest linkage disequilibrium values. Four of the 11 unique haplotypes identified in the HLA-homozygous cord blood units were the top-ranking haplotypes identified and were present in 18% of the cohort. This is the first study to report on HLA allele and haplotype frequencies for umbilical cord blood units from the Barcelona Cord Blood Bank and the largest study to date involving two fields of HLA resolution typing of Spanish registry data.
    MeSH term(s) 3' Untranslated Regions ; Alleles ; Blood Banks ; Female ; Fetal Blood/immunology ; Gene Frequency ; Genotype ; HLA-A Antigens/genetics ; HLA-A2 Antigen/genetics ; HLA-B Antigens/genetics ; HLA-B44 Antigen/genetics ; HLA-C Antigens/genetics ; HLA-DQ beta-Chains/genetics ; HLA-DRB1 Chains/genetics ; Haplotypes ; Homozygote ; Humans ; Linkage Disequilibrium ; Male ; Mothers ; Polymorphism, Single Nucleotide ; Pregnancy
    Chemical Substances 3' Untranslated Regions ; HLA-A Antigens ; HLA-A*02:01 antigen ; HLA-A2 Antigen ; HLA-A29 antigen ; HLA-B Antigens ; HLA-B*44:03 antigen ; HLA-B44 Antigen ; HLA-C Antigens ; HLA-C*04:09N antigen ; HLA-C*16 antigen ; HLA-DQ beta-Chains ; HLA-DQB1 antigen ; HLA-DRB1 Chains ; HLA-DRB1*07 antigen
    Language English
    Publishing date 2019-08-06
    Publishing country England
    Document type Journal Article
    ZDB-ID 2845111-9
    ISSN 2059-2310 ; 2059-2302
    ISSN (online) 2059-2310
    ISSN 2059-2302
    DOI 10.1111/tan.13644
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    Zs.A 661: Show issues Location:
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  8. Article ; Online: Development of an in-house bone marrow collection kit: The Catalan bone marrow transplantation group experience.

    Fernandez-Sojo, Jesus / Valdivia, Elena / Esquirol, Albert / Portos, Jose-Manuel / Rovira, Montse / Suarez, Maria / Diaz-de-Heredia, Cristina / Uría, Maria-Luz / Ortí, Guillermo / Ferra, Christelle / Mussetti, Alberto / Paviglianiti, Annalisa / Marsal, Julia / Badell, Isabel / Lozano, Miquel / Gomez, David / Azqueta, Carmen / Martorell, Lluis / Rubio, Nuria /
    Garcia-Buendia, Ana / Villa, Juliana / Carreras, Enric / Querol, Sergio

    Vox sanguinis

    2023  Volume 118, Issue 9, Page(s) 783–789

    Abstract: Background and objectives: Bone marrow (BM) harvesting is one of the essential sources of stem cells for haematopoietic stem cell transplantation. In 2019, commercial BM collection kits became unavailable in Europe. Consequently, we created an in-house ... ...

    Abstract Background and objectives: Bone marrow (BM) harvesting is one of the essential sources of stem cells for haematopoietic stem cell transplantation. In 2019, commercial BM collection kits became unavailable in Europe. Consequently, we created an in-house BM collection kit as an alternative.
    Materials and methods: We compared two groups of BM collections. The first collections were taken using an in-house kit from June 2022 through February 2023 and the second with a commercial kit from February 2021 through May 2022. These all took place at seven collection centres (CC). We analysed the harvest quality (cell blood count, CD34+ cells, viability, potency and sterility), the incidents occurring with each kit and the time to neutrophil and platelet engraftment in recipients.
    Results: A total of 23 donors underwent BM harvesting with the in-house kit and 23 with the commercial one. Both cohorts were comparable regarding donor characteristics, CC and time to procedure. No statistical differences were found in harvest quality between the in-house and commercial kits. A new transfusion set was required in three BM harvests (13%) with the in-house kit because of filter clogging. The median time to neutrophil and platelet engraftment was 21 days for both cohorts and 29 days (in-house) and 33 days (commercial), p = 0.284, respectively.
    Conclusion: The in-house BM collection kit offers a real approach to solve the diminished supply of commercial kits. A higher risk of filter clogging was observed compared with commercial kits due to the lack of 850 and 500 μm filters.
    MeSH term(s) Humans ; Bone Marrow Transplantation/methods ; Bone Marrow ; Hematopoietic Stem Cell Transplantation ; Transplantation, Homologous ; Tissue Donors
    Language English
    Publishing date 2023-08-02
    Publishing country England
    Document type Journal Article
    ZDB-ID 80313-3
    ISSN 1423-0410 ; 0042-9007
    ISSN (online) 1423-0410
    ISSN 0042-9007
    DOI 10.1111/vox.13499
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  9. Article ; Online: A hub-and-spoke model to deliver effective access to chimeric antigen receptor T-cell therapy in a public health network: the Catalan Blood and Tissue Bank experience.

    Fernandez-Sojo, Jesus / Delgadillo, Joaquim / Vives, Joaquim / Rodriguez, Luciano / Mendoza, Ana / Azqueta, Carmen / Garcia-Buendia, Ana / Valdivia, Elena / Martorell, Lluis / Rubio-Lopez, Nuria / Linares, Mónica / Alonso, Sofia / Ancochea, Agueda / García-Rey, Enric / García-Muñoz, Nadia / Medina, Laura / Querol, Sergio

    Cytotherapy

    2022  Volume 25, Issue 1, Page(s) 14–19

    Abstract: Background aims: To describe and analyze whether a hub-and-spoke organizational model could efficiently provide access to chimeric antigen receptor (CAR) T-cell therapy within a network of academic hospitals and address the growing demands of this ... ...

    Abstract Background aims: To describe and analyze whether a hub-and-spoke organizational model could efficiently provide access to chimeric antigen receptor (CAR) T-cell therapy within a network of academic hospitals and address the growing demands of this complex and specialized activity.
    Methods: The authors performed a retrospective evaluation of activity within the Catalan Blood and Tissue Bank network, which was established for hematopoietic stem cell transplantation to serve six CAR T-cell programs in academic hospitals of the Catalan Health Service. Procedures at six hospitals were followed from 2016 to 2021. Collection shipments of starting materials, CAR T-cell returns for storage and infusions for either clinical trials or commercial use were evaluated.
    Results: A total of 348 leukocytapheresis procedures were performed, 39% of which were delivered fresh and 61% of which were cryopreserved. The network was linked to seven advanced therapy medicinal product manufacturers. After production, 313 CAR T-cell products were shipped back to the central cryogenic medicine warehouse located in the hub. Of the units received, 90% were eventually administered to patients. A total of 281 patients were treated during this period, 45% in clinical trials and the rest with commercially available CAR T-cell therapies.
    Conclusions: A hub-and-spoke organizational model based on an existing hematopoietic stem cell transplantation program is efficient in incorporating CAR T-cell therapy into a public health hospital network. Rapid access and support of growing activity enabled 281 patients to receive CAR T cells during the study period.
    MeSH term(s) Humans ; Immunotherapy, Adoptive/methods ; Receptors, Chimeric Antigen ; Public Health ; Retrospective Studies ; Hematopoietic Stem Cell Transplantation ; Receptors, Antigen, T-Cell
    Chemical Substances Receptors, Chimeric Antigen ; Receptors, Antigen, T-Cell
    Language English
    Publishing date 2022-08-27
    Publishing country England
    Document type Journal Article ; Research Support, Non-U.S. Gov't
    ZDB-ID 2039821-9
    ISSN 1477-2566 ; 1465-3249
    ISSN (online) 1477-2566
    ISSN 1465-3249
    DOI 10.1016/j.jcyt.2022.07.011
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  10. Article ; Online: Post thawing viable CD34+ Cells dose is a better predictor of clinical outcome in lymphoma patients undergoing autologous stem cell transplantation.

    Fernandez-Sojo, Jesus / Cid, Joan / Azqueta, Carmen / Valdivia, Elena / Martorell, Lluis / Codinach, Margarita / Marsal, Julia / Mussetti, Alberto / Esquirol, Albert / Trabazo, Maria / Benitez, Maria Isabel / Ferra, Christelle / Fox, Maria Laura / Linares, Mónica / Alonso, Eva / García-Rey, Enric / García-Muñoz, Nadia / Medina, Laura / Castillo-Flores, Nerea /
    Vall-Llovera, Ferran / Garcia, Antoni / Pinacho, Asuncion / Talarn, Carme / Arroba, Jose Garcia / Coll, Rosa / Santos, Mireia / Valero, Oliver / Carreras, Enric / Lozano, Miquel / Querol, Sergio

    Bone marrow transplantation

    2022  Volume 57, Issue 8, Page(s) 1341–1343

    MeSH term(s) Antigens, CD34 ; Hematopoietic Stem Cell Transplantation ; Humans ; Lymphoma/therapy ; Peripheral Blood Stem Cell Transplantation ; Transplantation, Autologous
    Chemical Substances Antigens, CD34
    Language English
    Publishing date 2022-05-25
    Publishing country England
    Document type Letter
    ZDB-ID 632854-4
    ISSN 1476-5365 ; 0268-3369 ; 0951-3078
    ISSN (online) 1476-5365
    ISSN 0268-3369 ; 0951-3078
    DOI 10.1038/s41409-022-01722-6
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