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Ex vivo expansion

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https://www.readbyqxmd.com/read/28717408/sirt3-enhances-mesenchymal-stem-cell-longevity-and-differentiation
#1
Ryan A Denu
Mesenchymal stem cells (MSCs) are multipotent cells that are currently being investigated in a wide variety of clinical trials for their anti-inflammatory and immunomodulatory properties as well as their osteogenic and chondrogenic capabilities. However, there are considerable interdonor variability and heterogeneity of MSC populations, making it challenging to compare different products. Furthermore, proliferation, differentiation, and immunomodulation of MSCs decrease with aging and ex vivo expansion. The sirtuins have emerged as a class of protein deacylases involved in aging, oxidative stress, and metabolism...
2017: Oxidative Medicine and Cellular Longevity
https://www.readbyqxmd.com/read/28710530/proteomic-characterisation-reveals-active-wnt-signalling-by-human-multipotent-stromal-cells-as-a-key-regulator-of-beta-cell-survival-and-proliferation
#2
Miljan Kuljanin, Gillian I Bell, Stephen E Sherman, Gilles A Lajoie, David A Hess
AIMS/HYPOTHESIS: Novel strategies to stimulate the expansion of beta cell mass in situ are warranted for diabetes therapy. The aim of this study was to elucidate the secretome of human bone marrow (BM)-derived multipotent stromal cells (MSCs) with documented islet regenerative paracrine function. We hypothesised that regenerative MSCs will secrete a unique combination of protein factors that augment islet regeneration. METHODS: Human BM-derived MSCs were examined for glucose-lowering capacity after transplantation into streptozotocin-treated NOD/severe combined immunodeficiency (SCID) mice and segregated into samples with regenerative (MSC(R)) vs nonregenerative (MSC(NR)) capacity...
July 14, 2017: Diabetologia
https://www.readbyqxmd.com/read/28708802/characterization-and-expansion-of-autologous-gmp-ready-regulatory-t-cells-for-treg-based-cell-therapy-in-patients-with-ulcerative-colitis
#3
Caroline J Voskens, Anika Fischer, Susanne Roessner, Carmen Lorenz, Simon Hirschmann, Raja Atreya, Clemens Neufert, Imke Atreya, Markus F Neurath, Gerold Schuler
BACKGROUND: A local imbalance between regulatory (Treg) and effector T cells is believed to play a major role in gut-specific inflammation, including ulcerative colitis (UC). Restoration of this balance through an adoptive Treg transfer is an attractive new treatment approach in patients who are refractory to current standard therapies. It was our goal to develop a Good Manufacturing Practices (GMP)-conform protocol for expansion of UC Treg cells as a rational backbone for future studies on Treg therapy in UC...
August 2017: Inflammatory Bowel Diseases
https://www.readbyqxmd.com/read/28706234/investigation-of-brain-tissue-infiltration-by-medulloblastoma-cells-in-an-ex-vivo-model
#4
Anuja Neve, Karthiga Santhana Kumar, Dimitra Tripolitsioti, Michael A Grotzer, Martin Baumgartner
Medulloblastoma (MB) is a paediatric cancer of the cerebellum that can develop cerebellar and leptomeningeal metastases. Local brain tissue infiltration, the underlying cause of metastasis and relapse, remains unexplored. We developed a novel approach to investigate tissue infiltration of MB using organotypic cerebellum slice culture (OCSC). We show that cellular and structural components of cerebellar tissue in OCSCs are maintained for up to 30 days ex vivo, and that OCSCs foster tumour growth and cell proliferation...
July 13, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28701507/the-innate-cytokines-il-25-il-33-and-tslp-cooperate-in-the-induction-of-type-2-innate-lymphoid-cell-expansion-and-mucous-metaplasia-in-rhinovirus-infected-immature-mice
#5
Mingyuan Han, Charu Rajput, Jun Y Hong, Jing Lei, Joanna L Hinde, Qian Wu, J Kelley Bentley, Marc B Hershenson
Early-life respiratory viral infection is a risk factor for asthma development. Rhinovirus (RV) infection of 6-d-old mice, but not mature mice, causes mucous metaplasia and airway hyperresponsiveness that are associated with the expansion of lung type 2 innate lymphoid cells (ILC2s) and are dependent on IL-13 and the innate cytokine IL-25. However, contributions of the other innate cytokines, IL-33 and thymic stromal lymphopoietin (TSLP), to the observed asthma-like phenotype have not been examined. We reasoned that IL-33 and TSLP expression are also induced by RV infection in immature mice and are required for maximum ILC2 expansion and mucous metaplasia...
July 12, 2017: Journal of Immunology: Official Journal of the American Association of Immunologists
https://www.readbyqxmd.com/read/28700636/large-scale-ex-vivo-generation-of-human-neutrophils-from-cord-blood-cd34-cells
#6
Zhenwang Jie, Yu Zhang, Chen Wang, Bin Shen, Xin Guan, Zhihua Ren, Xinxin Ding, Wei Dai, Yongping Jiang
Conventional high-dose chemotherapy frequently leads to severe neutropenia, during which patients experience a high risk of infection. Although support care with donor's neutrophils is possible this choice is largely hampered by the limited availability of matched donors. To overcome this problem, we explored a large-scale ex vivo production of neutrophils from hematopoietic stem cells (HSCs) using a four-stage culture approach in a roller-bottle production platform. We expanded CD34+ HSCs isolated from umbilical cord blood (UCB) using our in-house special medium supplemented with cytokine cocktails and achieved about 49000-fold expansion of cells, among which about 61% were differentiated mature neutrophils...
2017: PloS One
https://www.readbyqxmd.com/read/28700319/adamts1-responds-to-systemic-cues-and-gates-adipogenesis
#7
Janica C Wong, Brian J Feldman
Intuitively, excess caloric intake causes adipose tissue expansion. However, the signals and mechanisms by which this systemic trigger directs a local response in the adipose tissue are incompletely understood. Both hypertrophy of existing adipocytes and the generation of new adipocytes through differentiation of adipocyte precursor cells (APCs), contribute to adipose tissue expansion in response to changes in the diet. Ex vivo studies of this process elucidated an elegant network of mostly transcription factors that drive APCs through the differentiation (adipogenesis) process...
July 10, 2017: Adipocyte
https://www.readbyqxmd.com/read/28698551/laminin-511-and-521-based-matrices-for-efficient-ex-vivo-expansion-of-human-limbal-epithelial-progenitor-cells
#8
Naresh Polisetti, Lydia Sorokin, Naoki Okumura, Noriko Koizumi, Shigeru Kinoshita, Friedrich E Kruse, Ursula Schlötzer-Schrehardt
Optimization of culture conditions for human limbal epithelial stem/progenitor cells (LEPC) that incorporate the in vivo cell-matrix interactions are essential to enhance LEPC ex vivo-expansion and transplantation efficiency. Here, we investigate the efficacy of laminin (LN) isoforms preferentially expressed in the limbal niche as culture matrices for epithelial tissue engineering. Analyses of expression patterns of LN chains in the human limbal niche provided evidence for enrichment of LN-α2, -α3, -α5, -β1, -β2, -β3, -γ1, -γ2 and -γ3 chains in the limbal basement membrane, with LN-α5 representing a signature component specifically produced by epithelial progenitor cells...
July 11, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28694026/nano-technology-based-carriers-for-nitrogen-containing-bisphosphonates-delivery-as-sensitisers-of-%C3%AE-%C3%AE-t-cells-for-anticancer-immunotherapy
#9
Naomi O Hodgins, Julie Tzu-Wen Wang, Khuloud T Al-Jamal
Nitrogen containing bisphosphonates (N-BPs) including zoledronate (ZOL) and alendronate (ALD) inhibit farnesyl diphosphate synthase, and have been shown to have a cytotoxic affect against cancer cells as a monotherapy and to also sensitise tumour cells to destruction by γδ T cells. γδ T cells are a subset of human T lymphocytes and have a diverse range of roles in the immune system including the recognition and destruction of cancer cells. This property of γδ T cells can be harnessed for use in cancer immunotherapy through in vivo expansion or the adoptive transfer of ex vivo activated γδ T cells...
July 7, 2017: Advanced Drug Delivery Reviews
https://www.readbyqxmd.com/read/28693285/characterization-of-%C3%AE-%C3%AE-t-cells-in-patients-with-non-small-cell-lung-cancer
#10
Yi Bao, Li Guo, Juanfen Mo
Systemic immune defects that are associated with disease progression exist in a variety of malignancies. γδ T cells are innate-like lymphocytes that do not require self-major histocompatibility complex-restricted priming. Ex vivo-expanded circulating γδ T cells exhibit promising antitumor activity and are a potential candidate for the treatment of various malignancies, including non-small cell lung cancer (NSCLC). In the present study, flow cytometry was used as a method to study the phenotypes and characteristics of γδ T cells...
July 2017: Oncology Letters
https://www.readbyqxmd.com/read/28688754/strategy-for-erythroid-differentiation-in-ex%C3%A2-vivo-cultures-lentiviral-genetic-modification-of-human-hematopoietic-stem-cells-with-erythropoietin-gene
#11
Luisina A Cappellino, Ricardo B Kratje, Marina Etcheverrigaray, Claudio C Prieto
If cultured in appropriate conditions, such as supplementing culture media with costly cytokines and growth factors, hematopoietic stem/progenitor cells (HSPCs) from different origins have shown to be an adequate source of erythroid cells. This requirement turns erythroid cells production into a complicated process to be scaled-up for future applications. The aim of our work was to genetically modify HSPCs with human erythropoietin (hEPO) sequence by lentiviral transgenesis in order for cells to secrete the hormone into the culture medium...
July 6, 2017: Journal of Bioscience and Bioengineering
https://www.readbyqxmd.com/read/28685587/3-dimensional-nano-fibre-scaffold-for-ex-vivo-expansion-of-cord-blood-haematopoietic-stem-cells
#12
Seyed Hadi Mousavi, Saeid Abroun, Masoud Soleimani, Seyed Javad Mowla
Cord blood (CB) haematopoietic stem cell (HSC) is an alternative source of HSC transplantation. The limited cell number greatly restricts their clinic-scale therapeutic applications. The objective of this study was an ex vivo expansion of CB HSCs in a new three-dimensional polycaprolactone nano-scaffold coated with fibronectin (FN). First, we isolated CB CD34+ cells and cultured 10 days in presence of growth factors. The evaluation was performed by qRT-PCR, flow cytometry and clonogenicity. 3D PCL nano-scaffold coated with FN produced significantly higher total nucleated cells and CD34+ cells (p < ...
July 7, 2017: Artificial Cells, Nanomedicine, and Biotechnology
https://www.readbyqxmd.com/read/28675676/polyclonal-regulatory-t-cell-therapy-for-control-of-inflammation-in-kidney-transplants
#13
Sindhu Chandran, Qizhi Tang, Minnie Sarwal, Zoltan G Laszik, Amy L Putnam, Karim Lee, Joey Leung, Vinh Nguyen, Tara Sigdel, Erica C Tavares, Joshua Y C Yang, Marc Hellerstein, Mark Fitch, Jeffrey A Bluestone, Flavio Vincenti
Early subclinical inflammation in kidney transplants is associated with later graft fibrosis and dysfunction. Regulatory T cells (Tregs) can reverse established inflammation in animal models. We conducted a pilot safety and feasibility trial of autologous Treg cell therapy in three kidney transplant recipients with subclinical inflammation noted on 6-month surveillance biopsies. Tregs were purified from peripheral blood and polyclonally expanded ex vivo using medium containing deuterated glucose to label the cells...
July 4, 2017: American Journal of Transplantation
https://www.readbyqxmd.com/read/28674799/forward-rnai-screens-in-human-hematopoietic-stem-cells
#14
Roman Galeev, Christine Karlsson, Aurélie Baudet, Jonas Larsson
Identifying the genes and pathways that regulate self-renewal and differentiation in somatic stem cells is a central goal in stem cell and cancer biology. Here, we describe a method for RNA interference (RNAi)-based screens combined with next-generation sequencing (NGS) in primary human hematopoietic stem and progenitor cells (HSPCs). These cells are suitable targets for complex, selection-based screens using pooled lentiviral short hairpin RNA (shRNA) libraries. The screening approach presented in this chapter is a promising tool to dissect regulatory mechanisms in hematopoietic stem cells (HSCs) and somatic stem cells in general, and may be particularly useful to identify gene targets and modifiers that can be further exploited in strategies for ex vivo stem cell expansion...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28668076/ex-vivo-expanded-natural-killer-cells-from-breast-cancer-patients-and-healthy-donors-are-highly-cytotoxic-against-breast-cancer-cell-lines-and-patient-derived-tumours
#15
Mira M Shenouda, Amy Gillgrass, Tina Nham, Richard Hogg, Amanda J Lee, Marianne V Chew, Mahsa Shafaei, Craig Aarts, Dean A Lee, John Hassell, Anita Bane, Sukhbinder Dhesy-Thind, Ali A Ashkar
BACKGROUND: Natural killer (NK) cells play a critical role in cancer immunosurveillance. Recent developments in NK cell ex-vivo expansion makes it possible to generate millions of activated NK cells from a small volume of peripheral blood. We tested the functionality of ex vivo expanded NK cells in vitro against breast cancer cell lines and in vivo using a xenograft mouse model. The study aim was to assess functionality and phenotype of expanded NK cells from breast cancer patients against breast cancer cell lines and autologous primary tumours...
July 1, 2017: Breast Cancer Research: BCR
https://www.readbyqxmd.com/read/28667272/adiponectin-is-an-endogenous-anti-fibrotic-mediator-and-therapeutic-target
#16
Roberta G Marangoni, Yuri Masui, Feng Fang, Benjamin Korman, Gabriel Lord, Junghwa Lee, Katja Lakota, Jun Wei, Philipp E Scherer, Laszlo Otvos, Toshimasa Yamauchi, Naoto Kubota, Takashi Kadowaki, Yoshihide Asano, Shinichi Sato, Warren G Tourtellotte, John Varga
Skin fibrosis in systemic sclerosis (SSc) is accompanied by attrition of dermal white adipose tissue (dWAT) and reduced levels of circulating adiponectin. Since adiponectin has potent regulatory effects on fibroblasts, we sought to assess adiponectin signaling in SSc skin biopsies, and evaluate fibrosis in mice with adiponectin gain- and loss-of-function mutations. Furthermore, we investigated the effects and mechanism of action of agonist peptides targeting adiponectin receptors in vitro and in vivo. We found that adiponectin pathway activity was significantly reduced in a subset of SSc skin biopsies...
June 30, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28660094/aqueous-deficient-dry-eye-syndrome-post-orbital-radiotherapy-a-10-year-retrospective-study
#17
Shubha Tiwari, Anusha Bhatt, Jayalaxmi Nagamodi, Mohammad Javed Ali, Hasnat Ali, Milind N Naik, Vijay Anand P Reddy, Geeta K Vemuganti
PURPOSE: Despite advances in orbital radiotherapy (XRT), a significant proportion of patients develop ophthalmic complication like dry eye syndrome (DES). The study evaluates the prevalence of aqueous deficient DES (ADDE) and lacrimal gland (LG) changes through histologic evaluation and ex-vivo expansion potential postorbital XRT. METHODS: With the approval of the institutional review board, medical records of patients who underwent orbital XRT as management protocol were reviewed for evidence of ADDE using DEWS (Dry Eye Workshop) 2007 criteria (n = 51)...
June 2017: Translational Vision Science & Technology
https://www.readbyqxmd.com/read/28659917/a-two-phase-expansion-protocol-combining-interleukin-il-15-and-il-21-improves-natural-killer-cell-proliferation-and-cytotoxicity-against-rhabdomyosarcoma
#18
Juliane Wagner, Viktoria Pfannenstiel, Anja Waldmann, Judith W J Bergs, Boris Brill, Sabine Huenecke, Thomas Klingebiel, Franz Rödel, Christian J Buchholz, Winfried S Wels, Peter Bader, Evelyn Ullrich
Rhabdomyosarcoma (RMS) is the most common soft tissue malignancy in children. Despite intensive research in recent decades the prognosis for patients with metastatic or relapsed diseases has hardly improved. New therapeutic concepts in anti-tumor therapy aim to modulate the patient's immune system to increase its aggressiveness or targeted effects toward tumor cells. Besides surgery, radiotherapy and chemotherapy, immune activation by direct application of cytokines, antibodies or adoptive cell therapy are promising approaches...
2017: Frontiers in Immunology
https://www.readbyqxmd.com/read/28644973/il-18-il-15-il-12-synergy-induces-elevated-and-prolonged-ifn-%C3%AE-production-by-ex-vivo-expanded-nk-cells-which-is-not-due-to-enhanced-stat4-activation
#19
Evan Lusty, Sophie M Poznanski, Karen Kwofie, Talveer S Mandur, Dean A Lee, Carl D Richards, Ali A Ashkar
The synergistic effect of IL-18/IL-15/IL-12 stimulation potently activates NK cells, inducing high levels of IFN-γ production. As a result of this potent stimulatory effect, NK cell pre-activation with IL-18/IL-15/IL-12 is being developed as a cancer immunotherapy. Ex vivo expansion of NK cells enables the efficient generation of large numbers of NK cells for wide-scale and repeated therapeutic use, and is thus an important source of NK cells for clinical application. However, the effects of IL-18/IL-15/IL-12 stimulation on ex vivo expanded NK cells have not yet been assessed...
June 20, 2017: Molecular Immunology
https://www.readbyqxmd.com/read/28642819/adoptive-cell-therapy-using-pd-1-myeloma-reactive-t-cells-eliminates-established-myeloma-in-mice
#20
Weiqing Jing, Jill A Gershan, Grace C Blitzer, Katie Palen, James Weber, Laura McOlash, Matthew Riese, Bryon D Johnson
BACKGROUND: Adoptive cellular therapy (ACT) with cancer antigen-reactive T cells following lymphodepletive pre-conditioning has emerged as a potentially curative therapy for patients with advanced cancers. However, identification and enrichment of appropriate T cell subsets for cancer eradication remains a major challenge for hematologic cancers. METHODS: PD-1(+) and PD-1(-) T cell subsets from myeloma-bearing mice were sorted and analyzed for myeloma reactivity in vitro...
2017: Journal for Immunotherapy of Cancer
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