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Ex vivo expansion

Miliotou N Androulla, Papadopoulou C Lefkothea
BACKGROUND: Cancer is one of the leading causes of death worldwide. Over the years, a number of conventional cytotoxic approaches for neoplastic diseases has been developed. However, due to their limited effectiveness in accordance with the heterogeneity of cancer cells, there is a constant search for therapeutic approaches with improved outcome, such as immunotherapy that utilizes and enhances the normal capacity of the patient's immune system. METHODS: Chimeric Antigen Receptor (CAR) T-cell therapy involves genetic modification of patient's autologous T-cells to express a CAR specific for a tumor antigen, following by ex vivo cell expansion and re-infusion back to the patient...
April 17, 2018: Current Pharmaceutical Biotechnology
Yufeng Wang, Yibo Zhang, Tiffany Hughes, Jianying Zhang, Michael A Caligiuri, Don M Benson, Jianhua Yu
PURPOSE: Daratumumab and its use in combination with other agents is becoming a new standard of care for treatment of multiple myeloma (MM). We mechanistically studied how daratumumab acts on NK cells. EXPERIMENTAL DESIGN: Quantities of NK cells in peripheral blood (PB) and/or bone marrow (BM) of MM patients or healthy donors were examined by flow cytometry. NK cell apoptosis and the associated mechanism were assessed by flow cytometry and immunoblotting. Patients' NK cells were expanded in vitro using feeder cells...
April 17, 2018: Clinical Cancer Research: An Official Journal of the American Association for Cancer Research
Yi Luo, Lijian Shao, Jianhui Chang, Wei Feng, Y Lucy Liu, Michele H Cottler-Fox, Peter D Emanuel, Martin Hauer-Jensen, Irwin D Bernstein, Lingbo Liu, Xing Chen, Jianfeng Zhou, Peter J Murray, Daohong Zhou
Uncovering the cellular and molecular mechanisms by which hematopoietic stem cell (HSC) self-renewal is regulated can lead to the development of new strategies for promoting ex vivo HSC expansion. Here, we report the discovery that alternative (M2)-polarized macrophages (M2-MΦs) promote, but classical (M1)-polarized macrophages (M1-MΦs) inhibit, the self-renewal and expansion of HSCs from mouse bone marrow (BM) in vitro. The opposite effects of M1-MΦs and M2-MΦs on mouse BM HSCs were attributed to their differential expression of nitric oxide synthase 2 (NOS2) and arginase 1 (Arg1), because genetic knockout of Nos2 and Arg1 or inhibition of these enzymes with a specific inhibitor abrogated the differential effects of M1-MΦs and M2-MΦs...
April 24, 2018: Blood Advances
Qiao-Chu Wang, Liang Zhen
OBJECTIVE: To establish a novel method for ex vivo expansion of natural killer cells from human umbilical blood, so as to provide the basis for NK cell therapy. METHODS: Mononucleated cells from human umbilical blood were harvested and suspended in a serum-free medium containing 5% autologous plasma, recombinant human IL-15 (50 ng/ml) and hydrocortisone sodium succinate (5×10-8 mol/L) at a concentration of 1.5×106 /ml, then the cells were seeded into flasks pre-coated with heparin sodium (100 U/cm2 ) or/and anti-human CD16 antibody (1 µg/cm2 )...
April 2018: Zhongguo Shi Yan Xue Ye Xue za Zhi
Xinglong Qu, Ying Tang, Shucheng Hua
The inflammation is the protective response of the body against various harmful stimuli; however, the aberrant and inappropriate activation tends to become harmful. The acute inflammatory response tends to resolved once the offending agent is subside but this acute response becomes chronic in nature when the body is unable to successfully neutralized the noxious stimuli. This chronic inflammatory microenvironment is associated with the release of various pro-inflammatory and oncogenic mediators such as nitric oxide (NO), cytokines [IL-1β, IL-2, interleukin-6 (IL-6), and tumor necrosis factor-α (TNF-α)], growth factor, and chemokines...
2018: Frontiers in Immunology
Naoyuki Chosa, Akira Ishisaki
Mesenchymal stem cells (MSCs) retain the ability to self-renew and differentiate into mesenchymal cells. Therefore, human MSCs are suitable candidates for use in regenerative medicine and cell therapies. Upon activation by tissue damage, MSCs contribute to tissue repair through a multitude of processes such as self-renewal, migration, and differentiation. However, loss of self-renewal and multi-lineage differentiation potential occurs at a high rate during cell doubling. Effective MSC therapies require the establishment of new techniques that preserve MSC multipotency after lengthy cell expansions...
February 2018: Japanese Dental Science Review
Earn Gan, Wendy Robson, Peter Murphy, Robert Pickard, Simon Pearce, Rachel Oldershaw
BACKGROUND: The highly plastic nature of adrenal cortex suggests the presence of adrenocortical stem cells (ACSC) but the exact in vivo identity of ACSC remains elusive. A few studies have demonstrated the differentiation of adipose or bone marrow-derived mesenchymal stem cells (MSC) into steroid-producing cells. We therefore investigated the isolation of multipotent MSC from human adrenal cortex. METHODS: Human adrenals were obtained as discarded surgical material...
April 5, 2018: Endocrine Connections
Paulo C M Urbano, Hans J P M Koenen, Irma Joosten, Xuehui He
A crucial issue for Treg-based immunotherapy is to maintain a bona fide Treg phenotype as well as suppressive function during and after ex vivo expansion. Several strategies have been applied to harness Treg lineage stability. For instance, CD28 superagonist stimulation in vitro , in the absence of CD3 ligation, is more efficient in promoting Treg proliferation, and prevention of pro-inflammatory cytokine expression, such as IL-17, as compared to CD3/CD28-stimulated Treg. Addition of the mTOR inhibitor rapamycin to Treg cultures enhances FOXP3 expression and Treg stability, but does impair proliferative capacity...
2018: Frontiers in Immunology
Hiroshi Kubota, Ralph L Brinster
Spermatogonial stem cells (SSCs) are the most primitive spermatogonia in the testis and have an essential role to maintain highly productive spermatogenesis by self-renewal and continuous generation of daughter spermatogonia that differentiate into spermatozoa, transmitting genetic information to the next generation. Since the 1950s, many experimental methods, including histology, immunostaining, whole-mount analyses, and pulse-chase labeling had been used in attempts to identify SSCs, but without success. In 1994, a spermatogonial transplantation method was reported that established a quantitative functional assay to identify SSCs by evaluating their ability to both self-renew and differentiate to spermatozoa...
March 30, 2018: Biology of Reproduction
Bin Guo, Xinxin Huang, Hal E Broxmeyer
PURPOSE OF REVIEW: Allogeneic hematopoietic cell transplantation (HCT) is a life-saving therapy for hematological and nonhematological diseases. Cord blood is a source of transplantable hematopoietic stem cells (HSCs), but limited numbers of HSCs in single cord blood units, which may cause delayed neutrophil, platelet, and immune cell reconstitution, is a major problem for efficient transplantation. Ex-vivo expansion and enhanced homing of cord blood HSC may overcome this disadvantage and improve its long-term engraftment...
March 31, 2018: Current Opinion in Hematology
Alessia Volpe, Francis Man, Lindsay Lim, Alex Khoshnevisan, Julia Blower, Philip J Blower, Gilbert O Fruhwirth
Metastasis is responsible for most cancer deaths. Despite extensive research, the mechanistic understanding of the complex processes governing metastasis remains incomplete. In vivo models are paramount for metastasis research, but require refinement. Tracking spontaneous metastasis by non-invasive in vivo imaging is now possible, but remains challenging as it requires long-time observation and high sensitivity. We describe a longitudinal combined radionuclide and fluorescence whole-body in vivo imaging approach for tracking tumor progression and spontaneous metastasis...
March 13, 2018: Journal of Visualized Experiments: JoVE
Pengfei Dong, David Prabhu, David L Wilson, Hiram G Bezerra, Linxia Gu
Stent deployment has been widely used to treat narrowed coronary artery. Its acute outcome in terms of stent under expansion and malapposition depends on the extent and shape of calcifications. However, no clear understanding as to how to quantify or categorize the impact of calcification. We have conducted ex vivo stenting characterized by the optical coherence tomography (OCT). The goal of this work is to capture the ex vivo stent deployment and quantify the effect of calcium morphology on the stenting. A three dimensional model of calcified plaque was reconstructed from ex vivo OCT images...
November 2017: International Mechanical Engineering Congress and Exposition: [proceedings]
Natalia Aparecida Nepomuceno, Karina Andrighetti de Oliveira-Braga, Liliane Moreira Ruiz, Aristides Tadeu Correia, Eduardo Zinoni Silva Pato, Luiz Fernando da Silva, Paulo Manuel Pêgo-Fernandes, Marcos Naoyuki Samano
BACKGROUND: Hemorrhagic shock-induced lung edema and inflammation are two of the main reasons for the rejection of lungs donated for transplantation. Hypertonic saline (HS) induces intravascular volume expansion and has considerable immunomodulating effects that might minimize edema. Our hypothesis is based on the use of a hypertonic solution for treatment of donors who are in shock in an attempt to increase the supply of lungs for transplantation. METHODS: A total of 80 rats were allocated to four groups: one group was given an infusion of normal saline (NS; n = 20), one group received HS; n = 20, a sham group (n = 20), and a Shock group (n = 20)...
May 2018: Journal of Surgical Research
Mitchell G Goldenberg, Jason Y Lee, Jethro Cc Kwong, Teodor P Grantcharov, Anthony Costello
OBJECTIVES: To systematically review and synthesize the validity evidence supporting intraoperative and simulation-based assessments of technical skill in urologic robotic-assisted surgery (RAS), and make evidence-based recommendations for the implementation of these assessments in urologic training. MATERIALS AND METHODS: A literature search of the MEDLINE, PsycINFO and Embase databases was performed. Articles using technical skill and simulation-based assessments in RAS were abstracted...
March 31, 2018: BJU International
Morand Piert, Prasad R Shankar, Jeffrey Montgomery, Lakshmi Priya Kunju, Virginia Rogers, Javed Siddiqui, Thekkelnaycke Rajendiran, Jason Hearn, Arvin George, Xia Shao, Matthew S Davenport
BACKGROUND: The study aims to assess the accuracy of multi-parametric prostate MRI (mpMRI) and18 F-choline PET/CT in tumor segmentation for clinically significant prostate cancer.18 F-choline PET/CT and 3 T mpMRI were performed in 10 prospective subjects prior to prostatectomy. All subjects had a single biopsy-confirmed focus of Gleason ≥ 3+4 cancer. Two radiologists (readers 1 and 2) determined tumor boundaries based on in vivo mpMRI sequences, with clinical and pathologic data available...
March 27, 2018: EJNMMI Research
Julian Lohmeyer, Thomas Nerreter, Julia Dotterweich, Hermann Einsele, Ruth Seggewiss-Bernhardt
Natural killer (NK) cells play a major role in host immunity against leukemia and lymphoma. However, clinical trials applying NK cells have not been as efficient as hoped for. Patients treated with RAF inhibitors exhibit increased tumor infiltration by immune cells suggesting that a combination of RAF inhibitors with immunotherapy might be beneficial. As MAPKs such as CRAF regulate NK cell functions, we performed an in vitro investigation on the potential of clinically relevant short acting tyrosine kinase inhibitors (TKIs) as potential adjuvants for NK cell therapy: NK cells from healthy human blood donors were thus treated with sorafenib, sunitinib or the pan-RAF inhibitor ZM336372 during ex vivo expansion...
March 24, 2018: Clinical and Experimental Immunology
Tor Paaske Utheim, Øygunn Aass Utheim, Panagiotis Salvanos, Catherine J Jackson, Stefan Schrader, Gerd Geerling, Amer Sehic
Limbal stem cell deficiency (LSCD) can result from a variety of corneal disorders, including chemical and thermal burns, infections, and autoimmune diseases. The symptoms of LSCD may include irritation, epiphora, blepharospasms, photophobia, pain, and decreased vision. There are a number of treatment options, ranging from nonsurgical treatments for mild LSCD to various forms of surgery that involve different cell types cultured on various substrates. Ex vivo expansion of limbal epithelial cells (LEC) involves the culture of LEC harvested either from the patient, a living relative, or a cadaver on a substrate in the laboratory...
March 23, 2018: Stem Cells Translational Medicine
Jae-Uk Jeong, Tung Nguyen Thanh Uong, Woong-Ki Chung, Taek-Keun Nam, Sung-Ja Ahn, Ju-Young Song, Sang-Ki Kim, Dong-Jun Shin, Eugene Cho, Kyoung Won Kim, Duck Cho, Mee Sun Yoon
BACKGROUND AIMS: Irradiation enhances the adhesion between natural killer (NK) cells and target cells by up-regulating intercellular adhesion molecule-1 (ICAM-1) on target cells. Therefore, we investigated the effect of irradiation-induced ICAM-1 expression on human cancer cells on NK cell-mediated cytotoxicity. METHODS: Expression levels of ICAM-1 on the target cell surface before and after irradiation of six human cancer cell lines (HL60, SKBR-3, T47D, HCT-116, U937 and U251) were analyzed by flow cytometry...
March 20, 2018: Cytotherapy
Lawrence J Liew, Linda Q Chen, Allen Y Wang, Magnus von Unge, Marcus Atlas, Rodney J Dilley
Epidermal cells with stem cell-like characteristics have been identified in the tympanic membrane (TM) and localized specifically to the umbo and annulus regions. While they have been proposed to play a role in the regeneration of both acute and chronic tympanic membrane perforations, evidence for the mechanism and regulation of their contribution is not yet described. Indeed, the behaviour of these putative stem cells is largely unknown, in part due to a lack of refined methods for efficient cell isolation...
March 23, 2018: Stem Cells and Development
Melania Lo Iacono, Eleonora Russo, Rita Anzalone, Elena Baiamonte, Giusi Alberti, Aldo Gerbino, Aurelio Maggio, Giampiero La Rocca, Santina Acuto
Wharton's jelly mesenchymal stromal cells (WJ-MSCs) have been recently exploited as a feeder layer in coculture systems to expand umbilical cord blood-hematopoietic stem/progenitor cells (UCB-HSPCs). Here, we investigated the role of WJ-MSCs in supporting ex vivo UCB-HSPC expansion either when cultured in direct contact (DC) with WJ-MSCs or separated by a transwell system or in the presence of WJ-MSC-conditioned medium. We found, in short-term culture, a greater degree of expansion of UCB-CD34+ cells in a DC system (15...
January 2018: Cell Transplantation
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