The washouts of discarded bone marrow collection bags and filters are a very abundant source of hMSCs

Capelli, Chiara; Salvadè, Agnese; Pedrini, Olga; Barbui, Valentina; Gotti, Elisa; Borleri, Gianmaria; Cabiati, Benedetta; Belotti, Daniela; Perseghin, Paolo; Bellavita, Piermario; Biondi, Andrea; Biagi, Ettore; Rambaldi, Alessandro; Golay, Joseé; Introna, Martino

doi:10.1080/14653240902960437

Cited by 26 publications

(32 citation statements)

References 20 publications

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“…MSCs were isolated and ex vivo expanded according to Good-ManufacturingPractice procedures (Cell-Therapy Laboratory "G. Lanzani", Ospedali Riuniti di Bergamo, authorization no. aM-189/2008 Agenzia Italiana del Farmaco, AIFA) (23,24). On day 7 after kidney transplant, autologous MSCs were administered intravenously (1.7 ϫ 10 6 cells and 2.0 ϫ 10 6 cells per kg body weight, respectively) after premedication with chlorphenamine and acetaminophen.…”

Section: Patientsmentioning

confidence: 99%

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Autologous Mesenchymal Stromal Cells and Kidney Transplantation

Perico

Casiraghi

Introna³

et al. 2011

Clinical Journal of the American Society of Nephrology

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SummaryBackground and objectives Mesenchymal stromal cells (MSCs) abrogate alloimmune response in vitro, suggesting a novel cell-based approach in transplantation. Moving this concept toward clinical application in organ transplantation should be critically assessed.Design, setting, participants & measurements A safety and clinical feasibility study (ClinicalTrials.gov, NCT00752479) of autologous MSC infusion was conducted in two recipients of kidneys from living-related donors. Patients were given T cell-depleting induction therapy and maintenance immunosuppression with cyclosporine and mycophenolate mofetil. On day 7 posttransplant, MSCs were administered intravenously. Clinical and immunomonitoring of MSC-treated patients was performed up to day 360 postsurgery.Results Serum creatinine levels increased 7 to 14 days after cell infusion in both MSC-treated patients. A graft biopsy in patient 2 excluded acute graft rejection, but showed a focal inflammatory infiltrate, mostly granulocytes. In patient 1 protocol biopsy at 1-year posttransplant showed a normal graft. Both MSCtreated patients are in good health with stable graft function. A progressive increase of the percentage of CD4 ϩ CD25 high FoxP3 ϩ CD127 Ϫ Treg and a marked inhibition of memory CD45RO ϩ RA Ϫ CD8 ϩ T cell expansion were observed posttransplant. Patient T cells showed a profound reduction of CD8 ϩ T cell activity. ConclusionsFindings from this study in the two patients show that MSC infusion in kidney transplant recipients is feasible, allows enlargement of Treg in the peripheral blood, and controls memory CD8 ϩ T cell function. Future clinical trials with MSCs to look with the greatest care for unwanted side effects is advised.

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Section: Patientsmentioning

confidence: 99%

“…MSCs were processed and cultured as previously reported (23,24). In brief, bone marrow aspirates were collected and nucleated cells were plated at 500,000 cells per cm 2 in minimum essential medium-␣ in the presence of 5% human platelet lysate.…”

Section: Msc Isolation and Expansionmentioning

confidence: 99%

Autologous Mesenchymal Stromal Cells and Kidney Transplantation

Perico

Casiraghi

Introna³

et al. 2011

Clinical Journal of the American Society of Nephrology

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282

287

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“…Reusable filters have smaller pore sizes (250 or 150µm) (11) . Dvorakova et al, Capelli et al, Sundin et al, and others reported successful isolation of hMSCs from bone marrow collection disposable sets, but no one has reported this from reusable filters (12)(13)(14) . In this study, we isolated and characterized hMSCs obtained from bone marrow collection filters of both disposable and reusable sets.…”

Section: Introductionmentioning

confidence: 99%

Isolamento e caracterização de células-tronco mesenquimais de filtros reutilizáveis e descartáveis de medula óssea

Deus

Normanton

Hamerschlak

et al. 2012

Einstein (São Paulo)

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OBJETIVO: Comparar as células-tronco mesenquimais humanas obtidas de filtros de coleta reutilizáveis àquelas coletadas em filtros descartáveis e caracterizá-las utilizando os critérios da International Society for Cellular Therapy. MÉTODOS: Foram isoladas células-tronco mesenquimais humanas de kits de coleta de medula óssea reutilizáveis e descartáveis, pela lavagem dos filtros com meio de cultura. As células isoladas foram caracterizadas de acordo com os critérios estabelecidos pela International Society for Cellular Therapy, por meio das técnicas de citometria de fluxo, diferenciação in vitro e citoquímica. RESULTADOS: As amostras foram obtidas de filtro descartável (n=3) e reutilizável (n=3). Todas as amostras obtidas de filtros descartáveis produziram células-tronco mesenquimais, e todas as células-tronco mesenquimais humanas derivadas de medula óssea preencheram os critérios estabelecidos pela International Society for Cellular Therapy. CONCLUSÃO: Este estudo mostrou que as células-tronco mesenquimais também podem ser obtidas de kits de coleta reutilizáveis (que permanecem em uso em vários centros, no mundo inteiro), para serem empregadas em pesquisa como uma fonte alternativa e ética.

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“…12 More precisely, 67 expansions were performed from 33 healthy donors, 4 β−thalassemia patients and 21 multiple sclerosis patients (Table 1). MSC were expanded from BM washouts or aspirates using human platelet lysate as previously described.…”

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confidence: 99%

“…MSC were expanded from BM washouts or aspirates using human platelet lysate as previously described. 12,13 Metaphases were prepared according to standard procedures 12 and analyzed by QFQ-banding. At least 20 metaphases per sample were analyzed.…”

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Frequent occurrence of non-malignant genetic alterations in clinical grade mesenchymal stromal cells expanded for cell therapy protocols

et al. 2014

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Human bone marrow mesenchymal stromal cells (BM-MSC) represent one of the most investigated "advanced therapeutic medicinal products".1 Recent safety concerns have focused attention on the possible malignant transformation due to mutations acquired during their large-scale in vitro expansion.2 Indeed, spontaneous oncogenic transformation has been described for murine MSC 3 although not for human cells, 4,5 with the exception of a few studies, 6which were subsequently retracted when it was realized that this was due to cross-contamination by a tumor cell line. 7,8 One single report has described the in vitro outgrowth of a transformed subpopulation from a normal BM sample.9 Furthermore, genetic aberrations of MSC have been very occasionally observed after long-term cultures 4,10,11 but interpreted to be related to senescence. 5In order to investigate the frequency of cytogenetic alterations in a broad "collection" of clinical-grade BM-MSC products, we performed cytogenetic analysis of 92 preparations expanded under Good Manufacturing Practice conditions.12 More precisely, 67 expansions were performed from 33 healthy donors, 4 β−thalassemia patients and 21 multiple sclerosis patients (Table 1). MSC were expanded from BM washouts or aspirates using human platelet lysate as previously described.12,13 Metaphases were prepared according to standard procedures 12 and analyzed by QFQ-banding. At least 20 metaphases per sample were analyzed. Karyotype was described according to the International System for Human Cytogenetic Nomenclature. Furthermore, p53 gene mutations were analyzed by deep sequencing of exons 5 to 11.Chromosomal abnormalities were detected in 17 of 86 expansions (19.8%). In all cases, the genetic lesions were spontaneous abnormalities.2 In 14 cases they were nonclonal: in 8 they involved one metaphase (MSC46, MSC70, MSC74, MSC79, MSC82, MSC87, MSC121, MSC126); in 5 two different chromosome abnormalities in two metaphases (MSC52, MSC55, MSC66 MSC100, MSC116). Only in one case were three different alterations in three metaphases found (MSC80). "Clonal chromosome changes" 2 were detected in 3 cases: in MSC114 monosomy of chromosome X was found in three metaphases, while in MSC119 and MSC122 inversion of chromosome 1 and a translocation involving chromosomes 9 and 4, respectively, were found in two metaphases. We also examined the results of multiple expansions from the same donors. Chromosomal anomalies were observed for 7 out of 13 donors (ns. 18, 20, 23, 32, 37, 50, 63), but these lesions were not recurrent and present only in some of the expansions performed. This suggests that chromosome aberrations do not associate with specific donors. In 6 cases, cytogenetic evaluation could not be performed on the final fresh P2 products due to lack of metaphases (Table 1) but in 5 of these the analyses could be repeated using a frozen P2 aliquot and in 4 cases karyotypes were normal. Similarly, when spontaneous and non-clonal abnormalities were detected, the karyotype analysis was repeated using a frozen P2 aliqu...

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The washouts of discarded bone marrow collection bags and filters are a very abundant source of hMSCs

Cited by 26 publications

References 20 publications

Autologous Mesenchymal Stromal Cells and Kidney Transplantation

Autologous Mesenchymal Stromal Cells and Kidney Transplantation

Isolamento e caracterização de células-tronco mesenquimais de filtros reutilizáveis e descartáveis de medula óssea

Frequent occurrence of non-malignant genetic alterations in clinical grade mesenchymal stromal cells expanded for cell therapy protocols

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