PubMed | Laboratoire Of Therapie Cellulaire, University of Cologne, Gustave Roussy Cancer Campus, CNRS Gustave Roussy Institute and 8 more.
Type: Journal Article | Journal: Oncoimmunology | Year: 2016
Dendritic cell-derived exosomes (Dex) are small extracellular vesicles secreted by viable dendritic cells. In the two phase-I trials that we conducted using the first generation of Dex (IFN--free) in end-stage cancer, we reported that Dex exerted natural killer (NK) cell effector functions in patients. A second generation of Dex (IFN--Dex) was manufactured with the aim of boosting NK and T cell immune responses. We carried out a phase II clinical trial testing the clinical benefit of IFN--Dex loaded with MHC class I- and class II-restricted cancer antigens as maintenance immunotherapy after induction chemotherapy in patients bearing inoperable non-small cell lung cancer (NSCLC) without tumor progression. The primary endpoint was to observe at least 50% of patients with progression-free survival (PFS) at 4 mo after chemotherapy cessation. Twenty-two patients received IFN--Dex. One patient exhibited a grade three hepatotoxicity. The median time to progression was 2.2 mo and median overall survival (OS) was 15 mo. Seven patients (32%) experienced stabilization of >4 mo. The primary endpoint was not reached. An increase in NKp30-dependent NK cell functions were evidenced in a fraction of these NSCLC patients presenting with defective NKp30 expression. Importantly, MHC class II expression levels of the final IFN--Dex product correlated with expression levels of the NKp30 ligand BAG6 on Dex, and with NKp30-dependent NK functions, the latter being associated with longer progression-free survival. This phase II trial confirmed the capacity of Dex to boost the NK cell arm of antitumor immunity in patients with advanced NSCLC.
Modalities for preparation, cryopreservation, thawing of hematopoietic stem cells and precautions for infusion to patient: Guidelines from the Francophone Society of Bone Marrow Transplantation and Cellular Therapy (SFGM-TC)
PubMed | Laboratoire Of Therapie Cellulaire, CHU Sart Tilman, Institute Paoli Calmettes, Nancy University Hospital Center and 8 more.
Type: Journal Article | Journal: Bulletin du cancer | Year: 2016
To date, despite an existing regulatory framework and standards, there are no true technical recommendations. A survey of 23cell processing facilities (France, Belgium and Switzerland) has allowed to overview current practices according to cellular products specifications upon arrival at the facility, with modalities for their preparation prior to cryopreservation, storage, thawing and finally for infusion to patient. Data analysis shows great variability of collected volumes and cell concentrations in cellular products. Despite homogeneous practices for handling cells at the facility, methods vary between centers, especially for the choice of cryoprotective solutions and thawing methods. During the workshop, practices have been discussed and summarized to write of recommendations about the following topics: processing and cryopreservation, thawing, bedside precautions (for infusion). This work identifies some improvements in terms of collection, choice of wash solution of thawed cells and validation of the conditions of carriage.
Chaput N.,French Institute of Health and Medical Research |
Chaput N.,Institute Gustave Roussy |
Chaput N.,Center dInvestigation Clinique en Biotherapie 507 |
Chaput N.,Laboratoire Of Therapie Cellulaire |
And 3 more authors.
Seminars in Immunopathology | Year: 2010
To communicate, cells are known to release in their environment proteins which bind to receptors on surrounding cells. But cells also secrete more complex structures, called membrane vesicles, composed of a lipid bilayer with inserted transmembrane proteins, enclosing an internal content of hydrophilic components. Exosomes represent a specific subclass of such secreted membrane vesicles, which, despite having been described more than 20 years ago by two groups studying reticulocyte maturation, have only recently received attention from the scientific community. This renewed interest originated first from the description of exosome secretion by antigen-presenting cells, suggesting a potential role in immune responses, and very recently by the identification of the presence of RNA (both messenger and microRNA) in exosomes, suggesting a potential transfer of genetic information between cells. In this review, we will describe the conclusions of 20 years of studies on the immune properties of exosomes and the most recent advances on their roles and potential uses as markers or as therapeutic tools during pathologies, especially in cancer. © 2010 Springer-Verlag.
Casteilla L.,Toulouse 1 University Capitole |
Casteilla L.,French National Center for Scientific Research |
Planat-Benard V.,Toulouse 1 University Capitole |
Planat-Benard V.,French National Center for Scientific Research |
And 6 more authors.
Current Stem Cell Research and Therapy | Year: 2010
Adipose tissue is the final tissue to develop and is strongly involved in energy homeostasis. It can represent up to 50% of body weight in obesity. Beside its metabolic role, endocrine functions appeared to play a key role in interconnecting adipose tissue with other tissues of the organism and in numerous physiological functions. The presence of adipocyte progenitors has long been demonstrated throughout life in the stromal fraction of adipose tissue. Now, it appears that these cells are multipotent and share numerous features with mesenchymal stem cells (MSC) derived from bone marrow. They also display some specificities and a strong pro-angiogenic potential. Altogether, these data emphasize the need to reconsider the potential of adipose tissue. Moreover, since fat pads are easy to sample, numerous and promising perspectives are now opening up in regenerative medicine, particularly in ischemic situations. © 2010 Bentham Science Publishers Ltd.
PubMed | Center atlantique, Laboratoire Of Therapie Cellulaire, Institute Paoli Calmettes, Nancy University Hospital Center and University Pierre and Marie Curie
Type: Journal Article | Journal: Pathologie-biologie | Year: 2014
Thousands of autologous and at less extent allogeneic hematopoietic stem cells (HSC) bags are cryopreserved in France. The majority of autologous HSC grafts are used within a year after collection. However, many bags are still unused and cryopreserved for many years. In France and on a European scale, the ever-growing number of cryopreserved bags represents a real economic health concern. Indeed, the cost of storage is about 100 per bag and per year. In addition, quality and therapeutic value of these long-term cryopreserved grafts needs to be evaluated. In the attempt to harmonize clinical practices between different French transplantation centers, the French Society of Bone Marrow Transplantation and Cell Therapies (SFGM-TC) set up its fourth annual series of workshops which brought together practitioners from its member centers across France. These workshops took place in September 2013 in Lille. In this article, we addressed the issue of the destruction of long-term cryopreserved grafts be them autologous or allogeneic and provide recommendations regarding their destruction.
Lavergne M.,ABCell Bio |
Lavergne M.,French Institute of Health and Medical Research |
Vanneaux V.,Laboratoire Of Therapie Cellulaire |
Delmau C.,French Institute of Health and Medical Research |
And 4 more authors.
Cell Proliferation | Year: 2011
Adult peripheral blood (PB) endothelial progenitor cells (EPC) are produced in the bone marrow and are able to integrate vascular structures in sites of neoangiogenesis. EPCs thus represent a potential therapeutic tool for ischaemic diseases. However, use of autologous EPCs in cell therapy is limited by their rarity in adult PB. Cord blood (CB) contains more EPCs than PB, and they are functional after expansion. They form primary colonies that give rise to secondary colonies, each yielding more than 10 7 cells after few passages. The number of endothelial cells obtained from one unit of CB is compatible with potential clinical application. EPC colonies can be securely produced, expanded and cryopreserved in close culture devices and endothelial cells produced in these conditions are functional as shown in different in vitro and in vivo assays. As CB EPC-derived endothelial cells would be allogeneic to patients, it would be of interest to prepare them from ready-existing CB banks. We show that not all frozen CB units from a CB bank are able to generate EPC colonies in culture, and when they do so, number of colonies is lower than that obtained with fresh CB units. However, endothelial cells derived from frozen CB have the same phenotypical and functional properties than those derived from fresh CB. This indicates that CB cryopreservation should be improved to preserve integrity of stem cells other than haematopoietic ones. Feasibility of using CB for clinical applications will be validated in porcine models of ischaemia. © 2011 Blackwell Publishing Ltd.
Maumus M.,French Institute of Health and Medical Research |
Peyrafitte J.-A.,University Paul Sabatier |
D'Angelo R.,French Institute of Health and Medical Research |
Fournier-Wirth C.,Laboratoire Of Recherche |
And 4 more authors.
International Journal of Obesity | Year: 2011
Objectives:Beside having roles in energy homeostasis and endocrine modulation, adipose tissue (AT) is now considered a promising source of mesenchymal stromal cells (adipose-derived stromal cells or ASCs) for regenerative medicine. Despite numerous studies on cultured ASCs, native human ASCs are rarely investigated. Indeed, the phenotype of ASCs in their native state, their localization within AT and comparison with bone marrow-derived mesenchymal stromal cells (BM-MSCs) has been poorly investigated.Design:To address these issues, the stroma vascular fraction (SVF) of human AT was extracted and native cell subtypes were isolated by immunoselection to study their clonogenic potential in culture. Immunohistology on samples of human AT in combination with reconstruction of confocal sections were performed in order to localize ASCs.Results:Compared with BM-MNCs, all native ASCs were found in the CD34 cell fraction of the AT-SVF. Native ASCs expressed classical mesenchymal markers described for BM-MSCs. Interestingly, CD34 expression decreased during ASC cell culture and was negatively correlated with cell proliferation rate. Immunohistological analysis revealed that native ASCs exhibited specific morphological features with protrusions. They were found scattered in AT stroma and did not express in vivo pericytic markers such as NG2, CD140b or alpha-smooth muscle actin, which appeared during the culture process. Finally, ASCs spontaneous commitment to adipocytic lineage was enhanced in AT from obese humans.Conclusions:The use of complementary methodological approaches to study native human ASCs revealed their immunophenotype, their specific morphology, their location within AT and their stemness. Furthermore, our data strongly suggest that human ASCs participate in adipogenesis during AT development. © 2011 Macmillan Publishers Limited All rights reserved.
Platelet lysate from whole blood-derived pooled platelet concentrates and apheresis-derived platelet concentrates for the isolation and expansion of human bone marrow mesenchymal stromal cells: Production process, content and identification of active components
Fekete N.,University of Ulm |
Gadelorge M.,Laboratoire Of Therapie Cellulaire |
Frst D.,University of Ulm |
Maurer C.,University of Ulm |
And 9 more authors.
Cytotherapy | Year: 2012
Background aims. The clinical use of human mesenchymal stromal cells (MSC) requires ex vivo expansion in media containing supplements such as fetal bovine serum or, alternatively, human platelet lysate (PL). Methods. Platelet concentrates were frozen, quarantine stored, thawed and sterile filtered to obtain PL. PL content and its effect on fibroblastcolony-forming unit (CFU-F) formation, MSC proliferation and large-scale expansion were studied. Results. PL contained high levels of basic fibroblast growth factor (bFGF), soluble CD40L (sCD40L), vascular cell adhesion molecule-1 (VCAM-1), intercellular adhesion molecule-1 (ICAM-1), platelet-derived growth factor AA (PDGF-AA), platelet-derived growth factor AB/BB (PDGF-AB/BB), chemokine (C-C) ligand 5 (CCL5; RANTES) transforming growth factor-β1 (TGF-β1) and chemokine (C-X-C) ligand 1/2/3 (GRO), with low batch-to-batch variability, and most were stable for up to 14 days. Inhibition of PDGF-BB and bFGF decreased MSC proliferation by about 20% and 50%, respectively. The strongest inhibition (about 75%) was observed with a combination of anti-bFGF + anti-PDGF-BB and anti-bFGF + anti-TGF-β1 + anti-PDGF-BB. Interestingly, various combinations of recombinant PDGF-BB, bFGF and TGF-β1 were not sufficient to promote cell proliferation. PL from whole blood-derived pooled platelet concentrates and apheresis platelet concentrates did not differ significantly in their growth-promoting activity on MSC. Conclusions. PL enhances MSC proliferation and can be regarded as a safe tool for MSC expansion for clinical purposes. \in particular, PDGF-BB and bFGF are essential components for the growth-promoting effect of PL, but are not sufficient for MSC proliferation. © 2012 Informa Healthcare.
Bourin P.,Laboratoire Of Therapie Cellulaire
Methods in molecular biology (Clifton, N.J.) | Year: 2011
Adipose tissue-derived stromal cells (ASCs) are promising tools for the new therapeutic field of regenerative medicine. Many research teams are intent on producing these cells for therapeutic purposes. The cell production must follow strict rules for safety and for constant quality of the cell product to ensure a reliable effect in patients. These rules are grouped under the generic term Good Manufacturing Practices. In this chapter, we describe the general concepts of ASC production for therapeutic use, explaining new terms such as traceability and qualification. We also introduce general requirements for the installation, equipment, material, and staff for the cell production. Then, we outline a general strategy for building a cell culture process. Finally, as an example, we describe the use of CellStack™ chambers and specific tube sets that allow for producing cells beginning with the stromal vascular fraction under near-closed conditions.
Boussion H.,groupe hospitalier Henri Mondor |
Chaput N.,French Institute of Health and Medical Research |
Chaput N.,Laboratoire Of Therapie Cellulaire
Oncologie | Year: 2015
Exosomes are nanovesicles released by a large number of cells in their physiological and pathological states. They have a specific composition and are formed from cytosolic proteins and nucleic acids surrounded by a complex lipid membrane. They play a significant role in intercellular exchanges and the regulation of immune responses. In tumour development, they are involved in angiogenesis, cell proliferation, the spread of metastases and in the regulation of cells responsible for anti-tumour immunity. They can be found within tumour cells and in the microenvironment cells, where they enable the metastatic niche to be created. In clinical practice with bronchial carcinoma, the use of exosomes has been developed not only as a diagnostic tool with “fluid biopsies” but also as a means of treatment, in particular reactivating the anti-tumour immune system. © 2015, Springer-Verlag France.