1University of Padova, Italy
2Venetian Institute of Molecular Medicine, Italy
3Great Ormond Street Institute of Child Health, University College of London, UK
Several acquired or congenital pathological conditions can affect skeletal muscle, leading to irreversible loss of muscle mass and function - volumetric muscle loss (VML). Decellularised tissues are natural scaffolds derived from tissues or organs, in which the cellular and nuclear contents are eliminated, and the tridimensional (3D) structure and composition of the extracellular matrix (ECM) are preserved. Such scaffolds retain biological activity, are biocompatible and do not show rejection after allogeneic or xenogeneic transplantation. Increasing reports suggest that decellularised tissues are promising candidates for their clinical application in patients affected by VML. We investigated the ability of three different decellularised skeletal muscle scaffolds to support muscle regeneration in axenogeneic immune-competent model of VML, in which the EDL muscle was surgically resected. All implanted acellular matrices, used to replace the resected muscles, were able to generate functional artificial muscles by promoting host myogenic cell migration and differentiation, as well as nervous fibres, vascular networks, and satellite cell (SC) homing. However, acellular tissue mainly composed of ECM allowed better myofibre 3D organization and the restoration of SC pool, when compared to scaffolds which also preserved muscular cytoskeletal structures. Finally, we showed that fibroblasts are indispensable to promote efficient migration and myogenesis by muscle stem cells across the scaffolds in vitro. This data strongly support the use of xenogenic acellular muscles as device to treat VML conditions in absence of donor cell implementation, as well as in vitro model for studying cell interplay during myogenesis.
Anna Urciuolo started her scientific career at University of Padova (Italy), in the lab of Prof. Paolo Bonaldo, where Anna Urciuolo had the possibility to reach high stages of competence on ECM, skeletal muscle homeostasis, disease and regeneration. By moving in the lab of Prof. Paolo De Coppi at UCL-ICH (UK) Anna Urciuolo entered in the field of tissue engineering, by applying decellularized organs for regenerative medicine. Then in Prof. Nicola Elvassoreʼs lab at University of Padova, Anna Urciuolo implement her specialization on tissue engineering, stem cells and biomaterials. At date, Anna Urciuolo a young principal investigator at the University of Padua.
1Anhanguera University – UNIDERP, Brazil
2Federal University of Mato Grosso do Sul, Brazil
Bothrops moojeni snake bite causes hemorrhage and myonecrosis, however the standard anti-venom therapy generally is ineffective in neutralizing the local effects. In this way, it is important to identify complementary therapies to the use of antivenom, such as medicinal plants. The objective was to compare the effectiveness of aerial parts of Sebastiania hispida and Byrsonima crassifolia to neutralize the muscle damage caused by B. moojeni. Wistar rats were used and was separated into 4 groups (n=4): Control (saline); Venom (Vbm); Venom and hydro-methanolic extract of S. hispida (VSh); Venom and water extract of B. crassifólia (VBc). The venom (40 µg/mL) or saline was injected in the gastrocnemius muscle of the right paw. The extracts were administered orally. After the periods of 24 hours, 3, 7 and 14 days, blood was collected for evaluation of creatine kinase (CK) and the gastrocnemius muscle was used for histological analysis. Vbm caused an increase in CK levels, indicating that there was muscle damage and only the VSh extract reduced CK levels. Muscle damage was confirmed by histological analysis, being observed necrosis, hemorrhage and edema in Vbm group. After 14 days the Vsh extract reduced hemorrhage, edema and necrosis, while Vcb only reduced the hemorrhage. Thus, medicinal plant extracts are great alternatives to counteract the local damage caused by B. moojeni and, among the two species analyzed, the extract of S. hispida was the most effective in neutralizing these effects.
Doroty Mesquita Dourado, PhD and postdoctoral studies from UNICAMP/SP. She works as a professor and researcher at University Anhanguera Uniderp in Campo Grande, MS. Has published more than 20 papers in reputed journals and has been serving as an editorial board member of repute. Their researches are carried out with medicinal plans of the Pantanal and Cerrado in the Laboratory of Toxinology and Medicinal Plants of the University Anhanguera Uniderp in Campo Grande, MS.
1Duke-NUS Medical School, National University of Singapore, Singapore
2Department of Surgery, National University of Singapore, Singapore
3Cardiovascular Research Institute, National University Heart Centre Singapore, Singapore
4BioLamina AB, Sweden
5National Heart Research Institute Singapore, National Heart Centre Singapore, Singapore
6National Heart Lung Institute, Imperial College London, UK
7Department of Clinical Sciences, Karolinska Institute and Karolinska University Hospital, Sweden
8University Medical Center Utrecht, The Netherlands
9Department of Medical Biochemistry and Biophysics, Karolinska Institute, Sweden
The human heart has limited regeneration capacity after birth and methods for safe and reproducible generation of stem cellderived therapy for use in patients have not been successful. These include culturing of the stem cells or its products in undefined conditions and on xenogenic materials like MatrigelTM that render the cells unsuitable for human therapy and extensive variations in the differentiation protocols. In order to replace the xenogenic MatrigelTM and the use of ROCK inhibitor for a fully defined protocol, we have explored the extracellular matrix (ECM) components in the heart and found that a cardiomyocyte laminin protein is a highly biologically culture matrix. Here, we show based on deep RNA sequencing human heart muscle, that laminin 221 (LN-221) is the most abundantly expressed laminin in the human heart. We synthesized LN-221 as a recombinant human protein and found it to drive pluripotent human embryonic stem cells (hESCs) to the cardiovascular lineage under fully defined human conditions. LN-221 induces specific biological effects in hESCs by down regulating genes involved in pluripotency and teratoma development, while up regulating genes for cardiac development. We have also identified a highly reproducible expression signatures during differentiation of two separate hESCs to cardiovascular progenitors (CVPs) that become beating cardiomyocytes(CMs). Cardiac transplantation of CVPs into ischemic reperfusion heart in farction region in mice resulted in the formation of human muscle bundles. These bundles were formed from single troponin negative CVPs which later organized itself in vivo into well-organized CMs with normal sarcomeres and gap junctions. Transplanted hearts also showed improved cardiac function by echocardiogram. Moving towards a clinically safe therapy, we investigated the safety of these CVPs using a teratoma assay and in vivo imaging. We propose that LN-221-mediated differentiation of hESCs to CVPs may be developed as a new and fully human methodology for regenerative cardiology.
Karl Tryggvason, MD, PhD is Professor at Duke-NUS Medical School, Singapore and Duke University, North Carolina, as well as Senior Professor at Karolinska Institutet in Stockholm. His research concerns the molecular nature, biology and diseases of basement membranes (BM), a special compartment of the extracellular matrix. His group has cloned almost all human BM proteins and clarified genetic causes of many BM-associated diseases, such as Alport and congenital nephrotic syndromes, junctional epidermolysis bullosa and congenital muscular dystrophy, as well as studied matrix metalloproteinases, including the discovery and crystal structure of MMP-2. His group has produced most laminins as recombinant human proteins and currently the group mainly studies how different laminin isoforms influence cell growth and stem cell differentiation. Tryggvason has published over 400 research articles. He is a member of the Finnish Academy of Sciences and the Swedish Royal Academy of Sciences, and has served for 18 years as a member of the Nobel Assembly and Committee for Physiology or Medicine at the Karolinska Institute. He has received several international awards, and he is co-founder of Bio Lamina AB, Stockholm, that produces laminins for cell biology and cell therapy purposes.
1Neurological Disorders Research Center, Hamad Bin Khalifa University, Qatar
2Weill Cornell Medical College-Qatar, Qatar
3Department of Virology, Cairo University, Qatar
Stress Granules (SGs) are dynamic ribonucleoprotein aggregates, which have been observed in cells subjected to environmental stresses, such as oxidative stress and heat shock (HS). Pluripotent stem cells (PSCs) are highly sensitive to oxidative stress, indicating the importance of SGs in regulating stem cell fate. In this study we compared the effects of oxidative (sodium arsenite (SA) and hydrogen peroxide (H2O2) and thermal HS) stressors on SG formation in human induced (hi) PSCs. The aim was to establish whether these granules have a role in regulating PSC self-renewal and differentiation. We found that SA and HS, but not H2O2, induce SG formation in hiPSCs. The analyses of these granules showed that they are canonical SGs, because (i) they contain the well-known SGs proteins (G3BP, TIAR, eIF4E, eIF4A, eIF3B, eIF4G, and PABP), (ii) they were found in juxtaposition to processing bodies (PBs), and (iii) they were disassembled after the removal of the stress. Consistent with the SG data, SA and HS, but not H2O2, promote eIF2α phosphorylation in hiPSCs forming SGs. An initial screening for pluripotent marker proteins recruited to SGs confirmed that LIN28A and L1TD1 were SG markers and identified DPPA5 as a novel pluripotent marker that was weakly recruited to SGs. Altogether; our data introduce new aspects of how hiPSCs respond to adverse environmental conditions.
Dr. Mohamed M Emara is a scientist at the Qatar Biomedical Research Institute and an Assistant Professor at Hamad Bin Khalifa University, Qatar. He received a PhD in Molecular Genetics and Biochemistry from Georgia State University in 2007 and did his postdoctoral studies at Harvard Medical School. Dr. Mohamed M Emara main research focus is to use human hiPSCs in neurological diseases modeling with a specialinterest on neurodevelopmental (ASD) and neurodegenerative diseases (PD). Another branch Dr. Mohamed M Emaraʼs lab is to understand the possible role of stress response program components in regulating stem cell self-renewal and differentiation, with special focus on neuronal differentiation.
1 Department of Biology, Southern University of Science and Technology, China
2Department of Biochemistry, Rush University Medical Center, USA
3Department of Pathology, University of Pittsburgh School of Medicine, USA
Precise control of mesenchymal stem cell (MSC) differentiation is critical for tissue development and regeneration. Our recent studies have shown that kindlin-2 is a key determinant of MSC fate decision. Depletion of kindlin-2 in MSCs is sufficient to induce adipogenesis and inhibit osteogenesis in vitro and in vivo. Mechanistically, kindlin-2 regulates MSC differentiation through controlling YAP1/TAZ at both the transcript and protein levels. Kindlin-2 physically associates with myosin light chain kinase (MLCK) in response to mechanical cues of cell microenvironment and intracellular signaling events and promotes myosin light chain phosphorylation. Loss of kindlin-2 inhibits RhoA activation and reduces myosin light chain phosphorylation, stress fiber formation and focal adhesion assembly, resulting in increased Ser127 phosphorylation, nuclear exclusion and ubiquitin ligase atrophin-1 interacting protein (AIP)4-mediated degradation of YAP1/TAZ. Our findings reveal a novel kindlin-2 signaling axis that senses the mechanical cues of cell microenvironment and controls MSC fate decision, and suggest a new strategy to regulate MSC differentiation, tissue repair and regeneration.
Dr. Chuanyue Wu is a professor and the Lombardi and Shinozuka Experimental Pathology Research Chair in the Department of Pathology, the University of Pittsburgh School of Medicine, USA. His research focuses on the molecular basis and functions of cell-extracellular matrix adhesion in regulation of cell behavior including cell differentiation, migration, proliferation and survival. He holds a B.S. and a M.S. from the East China University of Science and Technology in Shanghai, China and a Ph.D. from the University of Pittsburgh, Pittsburgh, PA, USA.
Revitacell Clinic, Spain
One of the most important and multifaceted diseases of modern society is the “metabolic syndrome”. This syndrome has not been fully understood, and for that reason there is still no effective treatment available. I suggest that a possible mechanism of stem cells is involved in the development of the metabolic syndrome. This point of view also allows us to consider other important pathologies that could have similar etiopathogenic pathways, such as aging. All these clinical situations could be the consequence of a progressive and persistent stem cell loss syndrome. The main result of this stem cell loss would be an irreversible fall of effective regenerative mesenchymal stem cell (MSC) groups. In this way, the normal body repair capabilities could become ineffective. This concept could open the possibility of a new treatment strategy in the metabolic syndrome and even aging: stem cell therapies.
Keywords: Metabolic Syndrome, Stem Cell, Aging
Miguel G Garber has over 34 years experience in Internal medicine and cardiology, with expertise in regenerative medicine, training and education, research, product development and senior management. He has more than 15 years working with Stem Cell, including building and managing the stem cell evaluation, explore and developing stem cell therapies for cardiomyopathies, osteoarthritis and regenerative medicine at Stem cell Therapeutics Department of American Medical Information Group and Clinica Quirurgica Quantum. He is currently serving as Medical Director of Regeners Clinic (International Regenerative Medicine), and Director of Clinica Castelló 68, Madrid, on going of several investigative researches involved Mesenchymal Stem Cells application (ASC) and Drug. Professor and clinical director of the Master in Regenerative Medicine, Dr. Garber has made a significant contribution to Stem cell Research. Actually he is involved in Adipose Stem Cell application. Umbilical cord Stem Cell Research and Regenerative Medicine open access journal, editorial board member. President of Spanish Society of Regenerative Medicine and Cell therapy (SEMERETEC).
Department of Neurosurgery, Dhaka Medical College & Hospital, Bangladesh
Purpose: Traumatic spinal cord injury (SCI) is a severe neurological disease which causes sensory loss, motor paralysis and autonomic difficulties which is usually treated with physiotherapy. Complete spinal cord injury with ASIA Grade A patients remains untreated and neglected in our country since long back. Here we describe an effective strategy for the treatment of complete spinal cord injury by autologous bone marrow derived mesenchymal stem cell therapy.
Material and Method: The study was conducted in the Neurosurgery Department, Dhaka Medical College Hospital, Dhaka Bangladesh from December 2015 to December 2017. Total 22 Patients with traumatic spinal cord injury (ASIA Grade-A) were included in this study. Total 60 ml of autologous bone marrow was aspirated and processed to prepare 6-7 ml of bone marrow aspirate concentrate (BMAC) which was transplanted at the site of cord injury. Bony alignment was done by decompression and stabilization. Post-surgical physiotherapy and regular follow up was given. Result: Surgical outcome was assessed by ASIA Grading. Among 22 patients, 10 patients (45.45%) improved by one grade, 7 patients (31.82%) improved by two grade, 2 patients (9.09%) improved by three grade, 3 patients (13.64%) did not show any improvement of any grade as because their follow-up period was short. 19 patients (86.36%) noticed sensory and 16 patients (72.72%) noticed autonomic improvement by some extent.
Conclusion: Stem cell therapy is safe and effective. Steady and focused progress in stem cell research will open the door for many disable patients in the country like Bangladesh.
Dr. Sukriti Das, Associate Professor, Department of Neurosurgery, Dhaka Medical College passed MBBS in 1992 from Mymensingh Medical College, Bangladesh. He also completed FCPS (General Surgery), FRCS Ed and MS (Neurosurgery). Currently he works on bone marrow derived stem cell therapy in the management of complete spinal cord injury.
1State Key Laboratory for the Diagnosis and Treatment of Infectious Diseases, Zhejiang University, China
2Collaborative Innovation Center for Diagnosis and Treatment of Infectious Diseases, China
3Chu Kochen Honors College, Zhejiang University, China
The immunomodulatory effects of mesenchymal stem cells (MSCs) in acute liver injury involve coordination with the entire immune system. But mouse liver immune cell at lasin acute liver injury and a system-wide approach that analyses of the interaction between all major lineages and MSCs on single-cell level has been poorly understood. To assess the interaction between the immune system and MSCs, we used 43 antibodies to analyze the immune cell compartment in carbon tetrachloride (CCl4)-injured mouse livers via mass cytometryon 1, 2, 3, and 7 days after MSC administration. We identified 18 immune cell subsets and found immune subsets in MSCs treated groups distinctly distinguished from control groups by high-dimensional analysis. Furthermore, MSCs can promote liver restitutive repair. It is closely related the suppressed proliferation of conventional natural killer (cNK) cells, plasma dendritic cells (pDCs), CD4+ T, CD8+ T, γδ T, and B cells and induced generation of monocytes, monocytederived macrophages, myeloid-derived suppressor cell (MDSC)-like cells and Tregs. Through high-dimensional analysis, we generated the landscape and dynamics of immune cell populations during CCl4-induced mouse liver injury and our results indicate that mass cytometry analysis of the entire immune system can identify immune subsets associated with MSCs, which might facilitate the development of MSC-based immune therapeutics for acute liver injury.
Keywords: Acute liver injury, immune cell landscape, mesenchymal stem cell, immunomodulatory, mass cytometry
Hongcui Cao is a professor from the State Key Laboratory for the Diagnosis and Treatment of Infectious Diseases, First Affiliated Hospital, College of Medicine at Zhejiang University, China.
1Federal University of Mato Grosso do Sul, Brazil
2Anhanguera University – UNIDERP, Brazil
Melanoma is considered one of the main current challenges of medicine. Herbal remedies used in traditional folk medicine provide a largely unexplored source of potential novel drugs. The aim of the study was investigated the in vitro and in vivo anticancer activity of leaves extract from S. Fluminensis on melanoma. Dried leaf samples were smashed and extracted with ethylic alcohol. Cytotoxicity tests were performed in NIH/3T3 (murine fibroblast) to determine Selectivity Index (IS) and on B16F10 (murine melanoma cells) where antitumor activity was expressed in GI50. In addition, BALB/c mice models were used to evaluate the in vivo anticancer activity. Mice were intraperitoneally injected with S. Fluminensis extract at doses of 100 and 200mg/kg on the 14th experimental day. The tumor inhibition ratio was determined after 24 days of treatment and the histopathological analyses of the tumor tissue and liver were compared. Analyzing the results, S. Fluminensis extract was active in B16F10 line (GI50: 4,37 µg/mL), which means that this value inhibited 50% of cell growth, been the extract considered a highly antineoplastic agent. The IS was 54, that is, the compound is 54-fold more active in B16F10 cells than in 3T3 cells. In the experimental model, the inhibition percentage of tumoral growth was 89,21% in the treated group with S. Fluminensis 200mg/kg and 33,97% in the 100mg/kg. Histopathology analysis of S. Fluminensis treated tumor tissue showed necrotic cells reduction, adipocytes presence, melanin deposition, vascularization and inflammatory process in a concentration dependent manner. On the liver, the animals treated with the extract on both concentrations showed normal hepatic organization, normal hepatocytes and absence of inflammatory focus. The results indicate that S. Fluminensis extract demonstrated both in vitro and in vivo anticancer activity, reducing the tumoral growth in B16F10 and could therefore be a highly antineoplastic agent.
Doroty Mesquita Dourado, PhD and postdoctoral studies from UNICAMP/SP. She works as a professor and researcher at University Anhanguera Uniderp in Campo Grande, MS. Has published more than 20 papers in reputed journals and has been serving as an editorial board member of repute. Their researches are carried out with medicinal plants of the Pantanal and Cerrado in the Laboratory of Toxinology and Medicinal Plants of the University Anhanguera Uniderp in Campo Grande, MS.
1State Key Laboratory for the Diagnosis and Treatment of Infectious Diseases, Zhejiang University, China
2Collaborative Innovation Center for Diagnosis and Treatment of Infectious Diseases, China
Background: The mechanisms of mesenchymal stem cell (MSC) transplantation to protect against acute liver injury have been well studied within the liver. However, the associated changes in the intestinal microbiota during this process are poorly understood.
Methods: In this study, compact bone-derived MSCs were injected into mice after carbon tetrachloride (CCl4) administration. Potential curative effect of MSC was evaluated by survival rate, biochemical and pathological results. Overall structural changes of microbial communities and alterations in the intestinal microbiota were assessed by sequenced 16S RNA amplicon libraries from the contents of the cecum and colon.
Results: MSCs significantly reduced the serum levels of aspartate transaminase and alanine transaminase and improved the histopathology and survival rate. Lower expression and discontinuous staining of zonulaoccludens, as well as disrupted tight junctions, were observed in CCl4-treated mice at 48 h compared with MSC-transplanted mice. Moreover, MSC transplantation led to intestinal microbiota changes that were reflected in the decreased abundance of Bacteroidetes S24-7 and Bacteroidaceae and increased abundance of Firmicutes Clostridiales, Rumino coccaceae, and Lacto bacillus at the initial time point compared with that in CCl4-treated mice. In addition, phylogenetic investigation of communities by the reconstruction of unobserved states (PICRUSt) based on the Green genes database revealed functional biomarkers of MSC-transplanted mice involved in cell motility, signal transduction, membrane transport, transcription and metabolism of lipids, cofactors, and vitamins, terpenoids and polyketides, as well as xenobiotics.
Conclusion: MSC infusion may be related to initial alterations on intestinal mucosal biology and homeostasis, which benefit liver repair.
Keywords: Mesenchymal stem cell, acute liver injury, intestinal, microbiota, microecology
University of Santiago de Compostela, Spain
Adenosine deaminases acting on RNA (ADARs) are the proteins responsible of the adenosine-to-inosine modification, critical for multiple RNA regulatory pathways. ADAR-mediated editing is associated with diseases ranging from neurological disorders to cancer. Even though ADAR1 has been recognized as an essential enzyme necessary for normal embryonic development, its function in pluripotency and reprogramming remains to be addressed. In this study we attempt to understand ADAR1 functions in somatic cell reprogramming. Here we show that, while being dispensable for self-renewal and pluripotency maintenance, ADAR1 editing activity is essential for the generation of induced pluripotent stem cells (iPSCs). Remarkably, we identified RNA editing-specific and stage-dependent functions of ADAR1 during reprogramming. Overall, our data links ADAR1 mediated A-to-I editing with the establishment of induced pluripotent stem cells and provides a better understanding of the reprogramming process.
Dr. Miguel Fidalgo obtained his Ph.D. in Molecular Medicine (2010) from University of Santiago de Compostela in Spain. He was a postdoctoral fellow with Dr. Jianlong Wang (2011-2016) at the Icahn School of Medicine at Mount Sinai, NY. Since 2016, he is a Principal Investigator at Center for Research in Molecular Medicine and Chronic Diseases (CiMUS), Spain. His lab is interested in understanding epigenetic regulation of transcription in the mammalian genome that governs cell fate decisions during embryonic development and human diseases. They aim to develop novel therapeutic strategies for age-related disorders as well as rare diseases.