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stem cell expansion

Asma Tahlawi, Michail E Klontzas, Mark C Allenby, José C F Morais, Nicki Panoskaltsis, Athanasios Mantalaris
Biomimetic materials are essential for the production of clinically relevant bone grafts for bone tissue engineering applications. Their ability to modulate stem cell proliferation and differentiation can be used to harness the regenerative potential of those cells and optimize the efficiency of engineered bone grafts. The arginyl-glycyl-aspartic acid (RGD) peptide has been recognized as the adhesion motif of various extracellular matrix proteins and can affect stem cell behaviour in biomaterials. Attempts to functionalize biomaterials with RGD have been limited to a maximum of 1-3 mm thickness scaffolds, overlooking the issue of core infiltration which represents a major hurdle in developing real thickness scaffolds...
December 11, 2018: Journal of Tissue Engineering and Regenerative Medicine
Jianting Shi, Chenyi Xue, Wen Liu, Hanrui Zhang
Macrophages play important roles in many diseases. We describe a protocol and the associated resources for the differentiation of human induced pluripotent stem cell-derived macrophages (IPSDM) and their applications in understanding human macrophage physiology and relevant diseases. The protocol uses an embryoid body-based approach with a combination of serum-free condition for hematopoiesis specification, followed by adherent culture with serum and M-CSF for myeloid expansion and macrophage maturation. The protocol produced an almost pure culture of CD45+ /CD18+ macrophages yielding up to 2 × 107 cells per 6-well plate of iPSCs within 24 days, demonstrating high efficiency, purity, and scalability...
December 10, 2018: Current Protocols in Stem Cell Biology
Atsunori Tsuchiya, Shuji Terai
Mouse fetal liver includes abundant hepatic stem/progenitor cells (HSPCs). Easy expansion with passage of HSPCs is necessary to obtain steady data. However, it is often difficult to enrich only HSPCs, and HSPCs can die when usual trypsin is used for replating. Here, we introduce serum-free long-term culture with passage of HSPCs using fetal mouse liver without a cell sorter.
2019: Methods in Molecular Biology
Dan Zhou, Lei Tan, Jian Li, Tanbin Liu, Yi Hu, Yalan Li, Sachiyo Kawamoto, Chengyu Liu, Shiyin Guo, Aibing Wang
Relative to the issues of off-target effects and the difficulty of inserting a long DNA fragment in the application of designer nucleases for genome editing, embryonic stem (ES) cell-based gene-targeting technology does not have these shortcomings and is widely used to modify animal/mouse genome ranging from large deletions/insertions to single nucleotide substitutions. Notably, identifying the relatively few homologous recombination (HR) events necessary to obtain desired ES clones is a key step, which demands accurate and reliable methods...
November 20, 2018: Journal of Visualized Experiments: JoVE
Jinning Ye, Kaitao Yuan, Weigang Dai, Kaiyu Sun, Guanghua Li, Min Tan, Wu Song, Yujie Yuan
Complement activation is associated with regional inflammation during acute gastrointestinal injury (AGI). This study is designed to explore how intracellular C3 activation in Paneth cells (PCs) affects regeneration of intestinal epithelium during AGI. AGI was induced in wildtype C57BL/6 mice, with sham operation employed as control. Exogenous C3 (1 mg, I.P.) was applied at 6 h post-surgery. Intestinal crypts harvested from ileum were cultured with presence or absence of C3 (20 μg/ml), with small interfering RNA against BST1 and complement activation inhibitor selectively applied in vitro...
December 7, 2018: International Immunopharmacology
Anna Barbara Di Stefano, Daniela Massihnia, Federica Grisafi, Marta Castiglia, Francesca Toia, Luigi Montesano, Antonio Russo, Francesco Moschella, Adriana Cordova
Angiogenesis is a crucial process for the maintenance of normal tissue physiology and it is involved in tissue remodeling and regeneration. This process is essential for adipose tissue maintenance. The adipose tissue is composed by different cell types including stromal vascular cells as well as adipose stem cells (ASCs). In particular, ASCs are multipotent somatic stem cells that are able to differentiate and secrete several growth factors; they are recently emerging as a new cell reservoir for novel therapies and strategies in many diseases...
November 30, 2018: European Journal of Cell Biology
Liping Guan, Xiaoyi Li, Jiali Wei, Zhihui Liang, Jing Yang, Xiufang Weng, Xiongwen Wu
BACKGROUND: As the implantation and long-term existence of tumor-specific T cells in host are the prerequisite for adoptive immunotherapy, memory stem T cells (TSCM ) with self-renewal and differentiation capacity show the greatest potential to implant and long-term exhibit function in vivo, compared with other T cells of differentiation stages. Hence, tumor-specific TSCM have become potential candidate for adoptive T cell therapy of cancer. Here, we reported a protocol to generate allogeneic antigen-specific CD8+ TSCM cells from human PBLs...
December 7, 2018: Stem Cell Research & Therapy
Tetsutaro Kikuchi, Masahiro Kino-Oka, Masanori Wada, Toyoshige Kobayashi, Midori Kato, Shizu Takeda, Hirotsugu Kubo, Tetsuya Ogawa, Hironobu Sunayama, Kazuhito Tanimoto, Manabu Mizutani, Tatsuya Shimizu, Teruo Okano
Introduction: Current production facilities for Cell-Based Health care Products (CBHPs), also referred as Advanced-Therapy Medicinal Products or Regenerative Medicine Products, are still dependent on manual work performed by skilled workers. A more robust, safer and efficient manufacturing system will be necessary to meet the expected expansion of this industrial field in the future. Thus, the 'flexible Modular Platform (fMP)' was newly designed to be a true "factory" utilizing the state-of-the-art technology to replace conventional "laboratory-like" manufacturing methods...
December 2018: Regenerative Therapy
Jessica Chang, Mee-Hae Kim, Eviryanti Agung, Sho Senda, Masahiro Kino-Oka
Introduction: Understanding how extracellular matrix (ECM) protein composition regulates the process of human induced pluripotent stem cell (hiPSC) colony formation may facilitate the design of optimal cell culture environments. In this study, we investigated the effect of migratory behaviors on hiPSC colony formation on various ECM-coated surfaces. Methods: To quantify how different ECM proteins affect migratory behavior during the colony formation process, single cells were seeded onto surfaces coated with varying concentrations of different ECM proteins...
June 2019: Regenerative Therapy
Valeriya V Solovyeva, Alisa A Shaimardanova, Daria S Chulpanova, Kristina V Kitaeva, Lisa Chakrabarti, Albert A Rizvanov
Tay-Sachs disease belongs to the group of autosomal-recessive lysosomal storage metabolic disorders. This disease is caused by β-hexosaminidase A (HexA) enzyme deficiency due to various mutations in α-subunit gene of this enzyme, resulting in GM2 ganglioside accumulation predominantly in lysosomes of nerve cells. Tay-Sachs disease is characterized by acute neurodegeneration preceded by activated microglia expansion, macrophage and astrocyte activation along with inflammatory mediator production. In most cases, the disease manifests itself during infancy, the "infantile form," which characterizes the most severe disorders of the nervous system...
2018: Frontiers in Physiology
Ge Ma, Fatima Abbasi, William T Koch, Howard Mostowski, Prajakta Varadkar, Brent Mccright
Neural stem cells (NSCs) isolated from a variety of sources are being developed as cellular therapies aimed at treating neurodegenerative diseases. During NSC culture and expansion it is important the cells do not differentiate prematurely because this may have an unfavorable effect on product quality and yield. In our study, we evaluated the use of Notch and Sox2 as markers for undifferentiated human and mouse NSCs. The expression of Notch2 and Sox2 during extensive-passage, low-oxygen culture and differentiation conditions were analyzed to confirm that the presence of these signature proteins directly correlates with the ability of NSCs to form new neurospheres and differentiate into multiple cell types...
December 2018: Cytotherapy
Nai Yang Fu, Bhupinder Pal, Yunshun Chen, Felicity C Jackling, Michael Milevskiy, François Vaillant, Bianca D Capaldo, Fusheng Guo, Kevin H Liu, Anne C Rios, Nicholas Lim, Andrew J Kueh, David M Virshup, Marco J Herold, Haley O Tucker, Gordon K Smyth, Geoffrey J Lindeman, Jane E Visvader
Long-lived quiescent mammary stem cells (MaSCs) are presumed to coordinate the dramatic expansion of ductal epithelium that occurs through the different phases of postnatal development, but little is known about the molecular regulators that underpin their activation. We show that ablation of the transcription factor Foxp1 in the mammary gland profoundly impairs ductal morphogenesis, resulting in a rudimentary tree throughout life. Foxp1-deficient glands were highly enriched for quiescent Tspan8hi MaSCs, which failed to become activated even in competitive transplantation assays, thus highlighting a cell-intrinsic defect...
December 3, 2018: Developmental Cell
Cristina L Esteves, F Xavier Donadeu
Although pericytes have long been known for their roles in blood vessel regulation, it was not until a decade ago that their tissue regeneration potential began to be considered, after studies showed that pericytes were the in vivo counterparts of mesenchymal stem/stromal cells (MSCs). The prospective isolation and culture expansion of pericytes brought great excitement as it opened the way to the therapeutic use of well-defined cell populations with known regenerative potential to overcome concerns associated with the use of traditional MSC preparations...
2018: Advances in Experimental Medicine and Biology
Anat Shemer, Jonathan Grozovski, Tuan Leng Tay, Jenhan Tao, Alon Volaski, Patrick Süß, Alberto Ardura-Fabregat, Mor Gross-Vered, Jung-Seok Kim, Eyal David, Louise Chappell-Maor, Lars Thielecke, Christopher K Glass, Kerstin Cornils, Marco Prinz, Steffen Jung
Microglia are yolk sac-derived macrophages residing in the parenchyma of brain and spinal cord, where they interact with neurons and other glial. After different conditioning paradigms and bone marrow (BM) or hematopoietic stem cell (HSC) transplantation, graft-derived cells seed the brain and persistently contribute to the parenchymal brain macrophage compartment. Here we establish that graft-derived macrophages acquire, over time, microglia characteristics, including ramified morphology, longevity, radio-resistance and clonal expansion...
December 6, 2018: Nature Communications
Jakub Suchánek, Klara Zoe Browne, Sherine Adel Nasry, Tereza Suchánková Kleplová, Nela Pilbauerová, Jan Schmidt, Tomáš Soukup
Recently, human natal dental pulp stem cells (hNDP-SCs) have been characterized in vitro and it has been shown that they satisfy criteria defining human mesenchymal stromal cells (MSCs), as proposed by the International Society for Cellular Therapy. However, these results were reached in the presence of xenogeneic expansion medium, which has the potential to alter the cells' functional capacity. To determine the validity of the previously reported hNDP-SCs characteristics for human cell therapy, we have cultured hNDP-SCs in allogeneic expansion medium...
September 2018: Brazilian Dental Journal
Mario Salvi, Danila Covelli
B cells have multiple actions on different phases of an immune reaction, mainly resulting in B and T cell-interaction (help), production of cytokines, regulation of dendritic cells and downregulation of regulatory B cells. The effectiveness of B cell depletion therapy is probably due to blockade of the antigen-presenting function of B cells, occurring very early in the setting of autoimmune reactions. B cells undergo a maturation process from stem cells during which the CD 20 antigen, which is the target of rituximab (RTX), is expressed from the stage of pre-B cells to mature and memory B cells, but not on plasma cells...
December 4, 2018: Eye
Walid Warda, Fabrice Larosa, Mathieu Neto Da Rocha, Rim Trad, Eric Deconinck, Ziad Fajloun, Cyril Faure, Denis Caillot, Marius Moldovan, Severine Valmary-Degano, Sabeha Biichle, Etienne Daguindau, Francine Garnache-Ottou, Sebastien Tabruyn, Olivier Adotévi, Marina Deschamps, Christophe Ferrand
Chronic myeloid leukemia (CML) is a chronic disease resulting in myeloid cell expansion through expression of the BCR-ABL1 fusion transcript. Tyrosine kinase inhibitors (TKI) have significantly increased survival of CML patients, and deep responders may consider stopping the treatment. However, more than 50% of patients relapse and restart TKI, subsequently suffering unknown toxicity. Because CML is a model immune system-sensitive disease, we hypothesize that chimeric antigen receptor (CAR) T cells targeting interleukin-1 receptor-associated protein (IL-1RAP) in quiescent CML stem cells may offer an opportunity for a permanent cure...
December 4, 2018: Cancer Research
Qianwen Wang, Lu Zhou, Tailing Wang, Xin Guo, Hao Yu, Jiaqi Wang
BACKGROUND: Rapid tissue expansion has been attempted, aiming at shortening the period of conventional expansion. However, it has scarcely been clinically applied because of its drawbacks such as low expansion efficiency and tissue destruction. Adipose-derived stem cell transplantation is a promising therapeutic method in regenerative medicine. However, its effects on rapid expansion remain poorly understood. METHODS: Twenty-four expanders were implanted in the dorsum of 12 pigs...
November 2018: Plastic and Reconstructive Surgery
Gamze Dereli Can, Özge Ekin Akdere, Mehmet Erol Can, Menemşe Gümüşderelioğlu
BACKGROUND: Several methods to cultivate limbal epithelial stem cells (LESCs) in vitro with the support of feeder layers and different growth medium formulations have been established for several years. The initial green medium consists of various ingredients that exhibit a non-optimal level of biosafety, therefore, different modifications have been made to suit it to safe clinical applications. However, the question of which formulation is the most appropriate remains to be answered...
November 30, 2018: Cytotherapy
Marcella La Noce, Francesca Paino, Luigi Mele, Gianpaolo Papaccio, Tarik Regad, Angela Lombardi, Federica Papaccio, Vincenzo Desiderio, Virginia Tirino
BACKGROUND: Cancer stem cells (CSCs) play a key role in cancer initiation, progression and chemoresistance. Epigenetic alterations have been identified as prominent factors that contribute to the CSCs phenotype. Here, we investigated the effects of the HDAC inhibitor valproic acid (VPA) and the demethylating agent, 5'azacytidine (DAC) on the stem phenotype of MG63 and Saos2 osteosarcoma cell lines. METHODS: Saos2 and MG63 cells were treated with DAC and VPA, alone and in combination...
December 3, 2018: Journal of Experimental & Clinical Cancer Research: CR
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