Sensebe L.,Etablissement Francais du Sang Center Atlantique |
Bourin P.,Etablissement Francais du Sang Pyrenees Mediterranee |
Tarte K.,French Institute of Health and Medical Research
Human Gene Therapy | Year: 2011
Because of their multi/pluripotency and immunosuppressive properties mesenchymal stem/stromal cells (MSCs) are important tools for treating immune disorders and for tissue repair. The increasing use of MSCs has led to production processes that need to be in accordance with Good Manufacturing Practice (GMP). In cellular therapy, safety remains one of the main concerns and refers to donor validation, choice of starting material, processes, and the controls used, not only at the batch release level but also during the development of processes. The culture processes should be reproducible, robust, and efficient. Moreover, they should be adapted to closed systems that are easy to use. Implementing controls during the manufacturing of clinical-grade MSCs is essential. The controls should ensure microbiological safety but also avoid potential side effects linked to genomic instability driving transformation and senescence or decrease of cell functions (immunoregulation, differentiation potential). In this rapidly evolving field, a new approach to controls is needed. © 2011 Mary Ann Liebert, Inc.
Mansuy J.-M.,Hopital Purpan |
Bendall R.,Royal Cornwall Hospital Trust |
Legrand-Abravanel F.,Hopital Purpan |
Legrand-Abravanel F.,French Institute of Health and Medical Research |
And 13 more authors.
Emerging Infectious Diseases | Year: 2011
Using a validated sensitive assay, we found hepatitis E virus (HEV) IgG in 52.5% of voluntary blood donors in southwestern France. This finding suggests HEV is highly endemic to this region. The high HEV prevalence may reflect local dietary practices, such as eating uncooked pork and game products.
Bigot N.,French Institute of Health and Medical Research |
Bigot N.,University of Rennes 1 |
Guerillon C.,French Institute of Health and Medical Research |
Guerillon C.,University of Rennes 1 |
And 12 more authors.
Cell Death and Disease | Year: 2015
Hypoxic niches help maintain mesenchymal stromal cell properties, and their amplification under hypoxia sustains their immature state. However, how MSCs maintain their genomic integrity in this context remains elusive, since hypoxia may prevent proper DNA repair by downregulating expression of BRCA1 and RAD51. Here, we find that the ING1b tumor suppressor accumulates in adipose-derived stromal cells (ADSCs) upon genotoxic stress, owing to SUMOylation on K193 that is mediated by the E3 small ubiquitin-like modifier (SUMO) ligase protein inhibitor of activated STAT protein B (PIAS4). We demonstrate that ING1b finely regulates the hypoxic response by triggering HIf1α proteasomal degradation. On the contrary, when mutated on its SUMOylation site, ING1b failed to efficiently decrease HIf1α levels. Consistently, we observed that the adipocyte differentiation, generally described to be downregulated by hypoxia, was highly dependent on ING1b expression, during the early days of this process. Accordingly, contrary to what was observed with HIf1α, the absence of ING1b impeded the adipogenic induction under hypoxic conditions. These data indicate that ING1b contributes to adipogenic induction in adipose-derived stromal cells, and thus hinders the phenotype maintenance of ADSCs. © 2015 Macmillan Publishers Limited All rights reserved.
PubMed | French Institute of Health and Medical Research, University of Rennes 1 and Etablissement Francais du Sang Pyrenees Mediterranee
Type: Journal Article | Journal: Stem cells (Dayton, Ohio) | Year: 2016
Long-term cultures under hypoxic conditions have been demonstrated to maintain the phenotype of mesenchymal stromal/stem cells (MSCs) and to prevent the emergence of senescence. According to several studies, hypoxia has frequently been reported to drive genomic instability in cancer cells and in MSCs by hindering the DNA damage response and DNA repair. Thus, we evaluated the occurrence of DNA damage and repair events during the ex vivo expansion of clinical-grade adipose-derived stromal cells (ADSCs) and bone marrow (BM)-derived MSCs cultured with platelet lysate under 21% (normoxia) or 1% (hypoxia) O2 conditions. Hypoxia did not impair cell survival after DNA damage, regardless of MSC origin. However, ADSCs, unlike BM-MSCs, displayed altered H2AX signaling and increased ubiquitylated H2AX levels under hypoxic conditions, indicating an impaired resolution of DNA damage-induced foci. Moreover, hypoxia specifically promoted BM-MSC DNA integrity, with increased Ku80, TP53BP1, BRCA1, and RAD51 expression levels and more efficient nonhomologous end joining and homologous recombination repair. We further observed that hypoxia favored mtDNA stability and maintenance of differentiation potential after genotoxic stress. We conclude that long-term cultures under 1% O2 were more suitable for BM-MSCs as suggested by improved genomic stability compared with ADSCs.
Gennero I.,University Paul Sabatier |
Gennero I.,Toulouse University Hospital Center |
Laurencin-Dalicieux S.,University Paul Sabatier |
Conte-Auriol F.,University Paul Sabatier |
And 17 more authors.
Bone | Year: 2011
Lysophosphatidic acid (LPA) is a lipid mediator that acts in paracrine systems via interaction with a subset of G protein-coupled receptors (GPCRs). LPA promotes cell growth and differentiation, and has been shown to be implicated in a variety of developmental and pathophysiological processes. At least 6 LPA GPCRs have been identified to date: LPA 1-LPA 6. Several studies have suggested that local production of LPA by tissues and cells contributes to paracrine regulation, and a complex interplay between LPA and its receptors, LPA 1 and LPA 4, is believed to be involved in the regulation of bone cell activity. In particular, LPA 1 may activate both osteoblasts and osteoclasts. However, its role has not as yet been examined with regard to the overall status of bone in vivo. We attempted to clarify this role by defining the bone phenotype of LPA 1 (-/-) mice. These mice demonstrated significant bone defects and low bone mass, indicating that LPA 1 plays an important role in osteogenesis. The LPA 1 (-/-) mice also presented growth and sternal and costal abnormalities, which highlights the specific roles of LPA 1 during bone development. Microcomputed tomography and histological analysis demonstrated osteoporosis in the trabecular and cortical bone of LPA 1 (-/-) mice. Finally, bone marrow mesenchymal progenitors from these mice displayed decreased osteoblastic differentiation. These results suggest that LPA 1 strongly influences bone development both qualitatively and quantitatively and that, in vivo, its absence results in decreased osteogenesis with no clear modification of osteoclasis. They open perspectives for a better understanding of the role of the LPA/LPA 1 paracrine pathway in bone pathophysiology. © 2011 Elsevier Inc.
Cappellesso-Fleury S.,Etablissement Francais du sang Pyrenees Mediterranee |
Puissant-Lubrano B.,University Paul Sabatier |
Puissant-Lubrano B.,Toulouse University Hospital Center |
Apoil P.-A.,University Paul Sabatier |
And 6 more authors.
Journal of Clinical Immunology | Year: 2010
Introduction: Bone marrow mesenchymal stem cells (BM-MSCs) and adipose tissue-derived stem cells share immunosuppressive capacities, suggesting that the latter could be a general property of stromal cells. Methods: To check this hypothesis, we compared human BM-MSC and fibroblasts for their in vitro multipotentiality, expandability and their immunomodulatory properties under normalized optimized culture conditions. Results: We report that, unlike BM-MSCs, fibroblasts cannot differentiate in vitro into adipocytes and osteoblasts and differ from BM-MSCs by the expression of membrane CD106, CD10 and CD26 and by the expression of collagen VII mRNA. Like BM-MSCs, fibroblasts are unable to provoke in vitro allogeneic reactions, but strongly suppress lymphocyte proliferation induced by allogeneic mixed lymphocyte culture (MLC) or mitogens. We show that fibroblasts' immunosuppressive capacity is independent from prostaglandin E2, IL-10 and the tryptophan catabolising enzyme indoleamine 2, 3-dioxygenase and is not abrogated after the depletion of CD8+ T lymphocytes, NK cells and monocytes. Conclusion: Finally, fibroblasts and BM-MSCs act at an early stage through blockage of lymphocyte activation, as demonstrated by down-regulation of GZMB (granzyme B) and IL2RA (CD25) expression. © Springer Science+Business Media, LLC 2010.
Daurat A.,University of Nimes |
Roger C.,University of Nimes |
Gris J.,University of Nimes |
Daurat G.,University of Nimes |
And 4 more authors.
Transfusion | Year: 2016
BACKGROUND Controversy exists regarding the safety of the different types of platelet (PLT) concentrates. This study was aimed at comparing the rate of adverse reactions associated with apheresis PLT concentrates (APCs) and pooled PLT concentrates (PPCs) both in donors and in recipients. STUDY DESIGN AND METHODS From the French national hemovigilance system, types and numbers of recipient adverse reactions were compared over a period from 2009 to 2011. Donor adverse reactions were available for 2010 and 2011. This study involved 23 of 26 French regions. Main outcomes were the rates of adverse reaction in recipients and serious adverse reaction in donors. RESULTS There were 790,854 PLT transfusions during the study period (477,747 [60%] with APCs, 313,107 [40%] with PPCs). APCs were associated with more adverse reactions (6244 vs. 2469 per 1,000,000, p < 0.001) and more severe and life-threatening reactions (respectively, 241 vs. 131 per 1,000,000, p < 0.001; and 182 vs. 121 per 1,000,000, p = 0.04). Mortality rates due to an adverse transfusion reaction were similar (15 vs. 6 per 1,000,000, p = 0.5). In donors, the number of whole blood (WB) donations was 4,722,685 whereas 266,095 apheresis procedures were performed. Serious adverse reactions were more frequent for apheresis procedures than for WB donations (5445 vs. 803 per 1,000,000, p < 0.001). CONCLUSION Our findings suggest that apheresis PLTs may be more hazardous than pooled PLTs both in recipients and in donors. This study calls for randomized trials to confirm or refute these results. © 2016 AABB.
PubMed | University of Nimes, McMaster University and Etablissement Francais du Sang Pyrenees Mediterranee
Type: Journal Article | Journal: Transfusion | Year: 2016
Controversy exists regarding the safety of the different types of platelet (PLT) concentrates. This study was aimed at comparing the rate of adverse reactions associated with apheresis PLT concentrates (APCs) and pooled PLT concentrates (PPCs) both in donors and in recipients.From the French national hemovigilance system, types and numbers of recipient adverse reactions were compared over a period from 2009 to 2011. Donor adverse reactions were available for 2010 and 2011. This study involved 23 of 26 French regions. Main outcomes were the rates of adverse reaction in recipients and serious adverse reaction in donors.There were 790,854 PLT transfusions during the study period (477,747 [60%] with APCs, 313,107 [40%] with PPCs). APCs were associated with more adverse reactions (6244 vs. 2469 per 1,000,000, p < 0.001) and more severe and life-threatening reactions (respectively, 241 vs. 131 per 1,000,000, p<0.001; and 182 vs. 121 per 1,000,000, p=0.04). Mortality rates due to an adverse transfusion reaction were similar (15 vs. 6 per 1,000,000, p = 0.5). In donors, the number of whole blood (WB) donations was 4,722,685 whereas 266,095 apheresis procedures were performed. Serious adverse reactions were more frequent for apheresis procedures than for WB donations (5445 vs. 803 per 1,000,000, p<0.001).Our findings suggest that apheresis PLTs may be more hazardous than pooled PLTs both in recipients and in donors. This study calls for randomized trials to confirm or refute these results.
Petrik J.,Scottish National Blood Transfusion Service |
Coste J.,Etablissement Francais du Sang Pyrenees Mediterranee |
Fournier-Wirth C.,Etablissement Francais du Sang Pyrenees Mediterranee
Transfusion and Apheresis Science | Year: 2011
Several miniaturized high throughput technologies have been developed in the last decade, primarily to study genomic structures and gene expression patterns under various conditions. At the same time, the microarrays, biosensors, integrated microfluidic lab-on-a-chip devices, next generation sequencing or digital PCR are gradually finding their diagnostic applications, although their suitability for specialised diagnostic fields has still to be assessed. In this review we discuss the potential applications of the new technologies to blood testing. © 2011 Elsevier Ltd.