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Ex vivo expansion

Mohamed B Ezzelarab
Regulatory T cells (Treg) are currently being evaluated in clinical allotransplantation for tolerance induction, with proven safety in humans with autoimmune diseases and graft-versus-host disease. A considerable amount of recent data suggests that additional factors may need to be validated, including the stability and commitment of newly discovered Treg subsets under inflammatory conditions, to further warrant safe and effective Treg-based therapeutic approaches. This review explores the opportunities and challenges of Treg-based cell therapy in xenotransplantation...
May 2018: Xenotransplantation
Katie Palen, Monica Thakar, Bryon D Johnson, Jill A Gershan
Adoptive cell therapy (ACT) of chimeric antigen receptor T cells has demonstrated remarkable success for the treatment of pediatric B-cell leukemia. For patients who are not candidates for chimeric antigen receptor T-cell therapy, ACT using tumor antigen-experienced polyclonal T cells may be a treatment option. Since leukemic blasts reside in the bone marrow and bone marrow is a preferred site for homeostatic proliferation of cytotoxic memory CD8 T cells, we hypothesized that bone marrow would be a source of activated T cells...
June 14, 2018: Journal of Pediatric Hematology/oncology
Hao Xu, Wenyue Cao, Liang Huang, Min Xiao, Yang Cao, Lei Zhao, Na Wang, Jianfeng Zhou
Chimeric antigen receptor T (CART) cell therapy has emerged as a potentially curative "drug" for cancer treatment. Cryopreservation of CART cells is necessary for their clinical application. Systematic studies on the effects of cryopreservation on the antitumor function of CART cells are lacking. Therefore, we compared the phenotypes and functions of CART cells that were cryopreserved during ex vivo expansion with those of freshly isolated populations. T cells expressing an anti-B-cell-maturation-antigen (BCMA) chimeric antigen receptor (CAR) were expanded in vitro for 10 days and then cryopreserved...
June 14, 2018: Cryobiology
Shukui Yu, Zhongxuan Li, Wei Zhang, Zhengde Du, Ke Liu, Denghua Yang, Shusheng Gong
Transplantation of bone marrow-derived endothelial progenitor cells (BM-EPCs) has been used as a therapeutic strategy for vascular repair. However, it remains controversial whether BM-EPCs exhibit clonal endothelial colony-forming cell (ECFC) capacity, a characteristic of true EPCs. The aim of this study was to isolate and explore the cellular properties of BM-ECFCs. We isolated BM-ECFCs from rat bone marrow with high purity via an optimized method. This approach involved the removal of selective colonies based on the conventional differential adhesive culture method used to isolate ECFCs from peripheral and umbilical cord blood...
June 13, 2018: Experimental Cell Research
Xiaojing Wang, Guowei Wang, Sarah Zingales, Baodong Zhao
Repairing bone defects poses a major orthopedic challenge because current treatments are constrained by the limited regenerative capacity of human bone tissue. Novel therapeutic strategies, such as stem cell therapy and tissue engineering, have the potential to enhance bone healing and regeneration, and hence may improve quality of life for millions of people. However, the ex vivo expansion of stem cells and their in vivo delivery pose technical difficulties that hamper clinical translation and commercial development...
June 13, 2018: Tissue Engineering. Part B, Reviews
Sandra P D'Angelo, Luca Melchiori, Melinda S Merchant, Donna B Bernstein, John Glod, Rosandra N Kaplan, Stephan A Grupp, William D Tap, Karen Chagin, Gwendolyn K Binder, Samik Basu, Daniel E Lowther, Ruoxi Wang, Natalie Bath, Alex Tipping, Gareth Betts, Indu Ramachandran, Jean-Marc Navenot, Hua Zhang, Daniel K Wells, Erin Van Winkle, Gabor Kari, Trupti Trivedi, Tom Holdich, Lini N Pandite, Rafael Amado, Crystal L Mackall
We evaluated safety and activity of autologous T cells expressing NY-ESO-1c259, an affinity-enhanced T cell receptor (TCR) recognizing an HLA-A2-restricted NY-ESO-1/LAGE-1a-derived peptide, in patients with metastatic synovial sarcoma (NY-ESO-1c259T cells). Confirmed antitumor responses occurred in 50% of patients (6/12) and were characterized by tumor shrinkage over several months. Circulating NY-ESO-1c259T cells were present post-infusion in all patients and persisted for at least 6 months in all responders...
June 11, 2018: Cancer Discovery
Clara T Nicolas, Raymond D Hickey, Kari L Allen, Zeji Du, Rebekah M Guthman, Robert A Kaiser, Bruce Amiot, Aditya Bansal, Mukesh K Pandey, Lukkana Suksanpaisan, Timothy R DeGrado, Scott L Nyberg, Joseph B Lillegard
BACKGROUND: Autologous hepatocyte transplantation after ex vivo gene therapy is an alternative to liver transplantation for metabolic liver disease. Here we evaluate ex vivo gene therapy followed by transplantation of single-cell or spheroid hepatocytes. METHODS: Pig and mouse hepatocytes were isolated, labeled with zirconium-89 and returned to the liver as single cells or spheroids. Biodistribution was evaluated through positron emission tomography-computed tomography...
June 5, 2018: Surgery
Mirasbek Kuterbekov, Paul Machillot, Pierre Lhuissier, Catherine Picart, Alain M Jonas, Karine Glinel
Porous polymeric microcarriers are a versatile class of biomaterial constructs with extensive use in drug delivery, cell culture and tissue engineering. Currently, most methods for their production require potentially toxic organic solvents with complex setups which limit their suitability for biomedical applications and their large-scale production. Herein, we report an organic, solvent-free method for the fabrication of porous poly(l-lactide) (PLLA) microcarriers. The method is based on the spherulitic crystallization of PLLA in its miscible blends with poly(ethylene glycol) (PEG)...
June 6, 2018: Acta Biomaterialia
Emanuela Corsini, Robert V House
One of the most powerful tools in immunotoxicology is the assessment of cytokines, the proteins/peptides responsible for regulating a variety of processes including immunity, inflammation, apoptosis, and hematopoiesis. Cytokine production measurements offer outstanding information and may eventually substitute for other more laborious procedures in the assessment of immunotoxicity. The particular profile of cytokine production provides indeed important information regarding the nature of many immunotoxic responses...
2018: Methods in Molecular Biology
Mehrnaz Safaee Talkhoncheh, Agatheeswaran Subramaniam, Mattias Magnusson, Praveen Kumar, Jonas Larsson, Aurélie Baudet
Despite extensive studies, it has been challenging to define culture conditions in which hematopoietic stem cells (HSCs) can be expanded ex vivo. Here, we show that chemical inhibition of the NF-κB signaling pathway leads to a significant improvement of HSC function from ex vivo cultured human umbilical cord blood derived CD34+ cells. We found a distinct peak of activation of the NF-κB pathway shortly after cells were put in culture, and consequently inhibition of the pathway was both necessary and sufficient during the first 24 hours of culture where it reduced the levels of several pro-inflammatory cytokines...
June 7, 2018: Haematologica
Lukas Capek, Cormac Flynn, Martin Molitor, Simon Chong, Petr Henys
OBJECTIVES: The technique of meshed skin grafting is known since 1960s. It was shown that there is a difference between the declared and real expansion ratio of the skin meshed graft. We hypothesize that the orientation of the Langer's lines in a split thickness skin graft is a key parameter in the resulting expansion ratio. METHODS: The skin graft meshing process was analyzed in two steps. In the first step, ex vivo uniaxial tests of human skin were performed. This served as an input for the constitutive model used for numerical simulations...
May 31, 2018: Burns: Journal of the International Society for Burn Injuries
Soraya Rasi Ghaemi, Bahman Delalat, Frances J Harding, Yazad D Irani, Keryn A Williams, Nicolas H Voelcker
Ex vivo-expanded buccal mucosal epithelial (BME) cell transplantation has been used to reconstruct the ocular surface. Methods for enrichment and maintenance of BME progenitor cells in ex vivo cultures may improve the outcome of BME cell transplantation. However, the parameter of cell seeding density in this context has largely been neglected. This study investigates how varying cell seeding density influences BME cell proliferation and differentiation on tissue culture polystyrene (TCPS). The highest cell proliferation activity was seen when cells were seeded at 5×104 cells/cm2 ...
January 1, 2018: Cell Transplantation
Andrea M Patterson, Louis M Pelus
Umbilical cord blood (UCB) is a highly valuable but low-quantity source of hematopoietic stem cells (HSCs) for life-saving transplantations. Recently in Nature Medicine, Guo et al. (2018) found that antagonism of a glycolysis-blocking pathway enhances ex vivo expansion of long-term HSCs from human UCB.
June 1, 2018: Cell Stem Cell
Véronique Lapostolle, Jean Chevaleyre, Pascale Duchez, Laura Rodriguez, Marija Vlaski-Lafarge, Ioanna Sandvig, Philippe Brunet de la Grange, Zoran Ivanovic
Feasibility of ex vivo expansion allows us to consider the steady-state peripheral blood as an alternative source of hematopoietic stem progenitor cells for transplantation when growth factor-induced cell mobilization is contraindicated or inapplicable. Ex vivo expansion dramatically enhances the in vivo reconstituting cell population from steady-state blood. In order to investigate phenotype and the expression of homing molecules, CD34, CD133, CD90, CD45RA, CD26 and CD9 expression was determined on sorted CD34+ cells according to CXCR4 (neg, low, bright) and CD133 expression before and after ex vivo expansion...
June 1, 2018: Haematologica
Jeffrey Barminko, Brad M Reinholt, Alexander Emmanuelli, Alannah N Lejeune, Margaret H Baron
The pathways that regulate the growth of erythroid progenitors are incompletely understood. In a computational analysis of gene expression changes during erythroid ontogeny, the vitamin D receptor ( Vdr ) nuclear hormone receptor transcription factor gene was identified in fetal and adult stages, but not at the embryonic stage of development. Vdr was expressed in definitive erythroid (EryD) progenitors and was downregulated during their maturation. Activation of Vdr signaling by the vitamin D3 agonist calcitriol increased the outgrowth of EryD colonies from fetal liver and adult bone marrow, maintained progenitor potential, and delayed erythroid maturation, as revealed by clonogenic assays, suspension culture, cell surface phenotype, and gene expression analyses...
June 12, 2018: Blood Advances
Paul Mayor, Kristen Starbuck, Emese Zsiros
During the last decade, the field of cancer immunotherapy has been entirely transformed by the development of new and more effective treatment modalities with impressive response rates and the prospect of long survival. One of the major breakthroughs is adoptive cell transfer (ACT) based on autologous T cells derived from tumor-infiltrating lymphocytes (TILs). TIL-based ACT is a highly personalized cancer treatment. T cells are harvested from autologous fresh tumor tissues, and after ex vivo activation and extensive expansion, are reinfused to patients...
May 23, 2018: Gynecologic Oncology
Philippe Hernigou, Gilles Guerin, Yasuhiro Homma, Arnaud Dubory, Nathalie Chevallier, Hélène Rouard, Charles Henri Flouzat Lachaniette
PURPOSE: Despite multiple possible treatments, the risk of collapse remains the main problem of osteonecrosis. Heart failure (HF). In an effort to address the reverse this issue, curative strategies with regenerative medicine are increasingly being considered. The aim of this technology is to halt or reverse progression of the disease to collapse. MATERIAL AND METHODS: The pioneering report by Hernigou published in 2002 was the first pilot study suggesting that injection of bone marrow stem cells was a safe approach able to improve osteonecrosis in patients with early stages...
May 24, 2018: International Orthopaedics
Matthias Aurich, Gunther O Hofmann, Florian Gras, Bernd Rolauffs
BACKGROUND: Autologous chondrocyte implantation (ACI) is a therapy for articular cartilage and osteochondral lesions that relies on notch- or trochlea-derived primary chondrocytes. An alternative cell source for ACI could be osteochondritis dissecans (OCD) fragment-derived chondrocytes. Assessing the potential of these cells, we investigated their characteristics ex vivo and after monolayer expansion, as monolayer expansion is an integral step of ACI. However, as monolayer expansion can induce de-differentiation, we asked whether monolayer-induced de-differentiation can be reverted through successive alginate bead culture...
May 24, 2018: BMC Musculoskeletal Disorders
Yixuan Lin, Kenward Vong, Koji Matsuoka, Katsunori Tanaka
Previously, Au(III) complexes coordinated with 2-benzoylpyridine ligand, BPy-Au, were prebound to a protein and used to discover a novel protein-directed labelling approach with propargyl ester functional groups. In this work, further examination discovered that gold catalysts devoid of the 2-benzoylpyridine ligand (ex/ NaAuCl4) had significantly reduced levels of protein labeling. Mechanistic investigations then revealed that BPy-Au and propargyl esters undergo a rare example of C(sp2)-C(sp) aryl-alkynyl cross coupling, likely via spontaneous reductive elimination...
May 23, 2018: Chemistry: a European Journal
Tito Panciera, Luca Azzolin, Daniele Di Biagio, Antonio Totaro, Michelangelo Cordenonsi, Stefano Piccolo
Here we present protocols to isolate primary differentiated cells and turn them into stem/progenitor cells (SCs) of the same lineage by transient expression of the transcription factor YAP. With this method, luminal differentiated (LD) cells of the mouse mammary gland are converted into cells that exhibit molecular and functional properties of mammary SCs. YAP also turns fully differentiated pancreatic exocrine cells into pancreatic duct-like progenitors. Similarly, to endogenous, natural SCs, YAP-induced stem-like cells ("ySCs") can be eventually expanded as organoid cultures long term in vitro, without further need of ectopic YAP/TAZ, as ySCs are endowed with a heritable self-renewing SC-like state...
May 7, 2018: Journal of Visualized Experiments: JoVE
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