| 1. |
- Banerjee, Debashish, et al.
(författare)
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Epithelial MUC1/Muc1 Promotes Cell migration, Reduces Apoptosis and Affects Levels of Mucosal Modulators During Acetylsalicylic Acid (Aspirin) Induced Gastropathy.
- 2015
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Ingår i: The Biochemical journal. - 1470-8728. ; 465:3, s. 423-431
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Tidskriftsartikel (refereegranskat)abstract
- MUC1 is a transmembrane mucin highly expressed in the stomach. While extensive research has uncovered many of its roles in cancer, knowledge about the functions of MUC1 in normal tissues is limited. Here we showed that acetylsalicylic acid (Aspirin, ASA) upregulated MUC1/Muc1 expression in the gastric mucosa of humans and wild type mice. ASA induced mucosal injury in all mice to a similar extent, however wild type animals and those chimeras with Muc1 on the epithelia recovered faster than Muc1 knock-out mice and chimeras carrying Muc1 on haematopoietic but not epithelial cells. MUC1 enhanced proliferation and migration of the human gastric cell line MKN-7, and increased resistance to apoptosis. The repeated treatment regime used caused a reduction in cyclooxygenase-1 expression, though wild type animals returned faster towards pre-treatment levels, and had increased cyclooxygenase-2 and vascular endothelial growth factor levels during recovery. Thus, we found that epithelial Muc1 is more important for the healing process than haematopoetic Muc1, and Muc1/MUC1 facilitates wound healing by enhancing cell migration and proliferation, protecting against apoptosis and mediating expression of mucosal modulators. Thus, MUC1 plays essential roles during wound healing, and development of treatment modalities targeting enhanced expression of MUC1 may be beneficial to treat mucosal wounds.
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| 2. |
- Berg, Malin, 1976, et al.
(författare)
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Replacement of a Tracheal Stenosis with a Tissue-Engineered Human Trachea Using Autologous Stem Cells: A Case Report
- 2014
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Ingår i: Tissue Engineering. Part A. - 1937-3341 .- 1937-335X. ; 20:1-2, s. 389-397
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Tidskriftsartikel (refereegranskat)abstract
- Cell-based therapies, involving tissue engineering represent interesting and potentially important strategies for treatment of patients with various disorders. Here, using a detergent-enzymatic method we prepared an intact 3-dimensional scaffold of an extracellular matrix (ECM) derived from a human cadaver donor trachea, which we repopulated with autologous stem cells and implanted into a 76-year old patient with tracheal stenosis including lower part of the larynx. Although the graft provided the patient with an open airway, a week after surgery, the mucous membrane of the graft was covered by a 1-2mm thick fungal infection, which was treated with local and systemic anti-fungal therapy. The airway lumen was postoperatively controlled by fiberbrandoscopy and found stable and sufficient. However, twenty-three days later the patient died due to cardiac arrest but with a patent, open, stable tracheal transplant and intact anastomoses. Histopathological results of the transplanted tracheal graft at autopsy showed a squamous but not ciliated epithelium, neovascularization, bundles of -sma positive muscle cells, serous glands and nerve fibres with S-100 positive nerve cells in the submucosa and intact chondrocytes in the cartilage. Our findings suggest that although autologous stem cells- engineered tracheal matrices may represent a tool for clinical tracheal replacement. Further preclinical studies are required for generating functional airway grafts and long term effects of such grafts.
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| 3. |
- Chougule, Priti, et al.
(författare)
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Isolation and characterization of human primary enterocytes from small intestine using a novel method.
- 2012
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Ingår i: Scandinavian journal of gastroenterology. - : Informa UK Limited. - 1502-7708 .- 0036-5521. ; 47:11, s. 1334-43
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Tidskriftsartikel (refereegranskat)abstract
- Abstract Cell culture studies of enterocytes are important in many fields. However, there are difficulties in obtaining cell lines from adult human intestine, such as microbial contamination of cultures from the tissue samples, short life span of enterocytes, overgrowth of mesenchymal cells, etc. Various model used to obtain adult intestinal cell lines are very complex requiring use of feeder layer or gel matrices. The aim of this study was to establish a novel method for the simple and reproducible isolation of human enterocytes. Enterocytes were isolated from SI samples (n = 5) obtained from cadaveric donors using a mechanical procedure, and separation with immunomagnetic beads coated with anti-EpCAM antibodies. Light and electron microscopy, flow cytometry and immunocytochemistry techniques were used to characterize the isolated cells. Immunohistochemical staining of normal SB biopsies confirmed that the cell cultures maintained an in vivo phenotype as reflected in cytokeratin expression CK18, CK20 and expression of intestine-specific markers such as sucrase isomaltase and maltase glucoamylase. Furthermore, the cells strongly expressed TLR-5, 6, 7, 8 and 10 and several molecules such as CD40, CD86, CD44, ICAM-1 and HLA-DR which are important in triggering cell-mediated immune responses. This novel technique provides a unique in vitro system to study the biology of enterocytes in normal conditions as well as to study inflammatory processes in various small bowel disorders.
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| 4. |
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| 5. |
- Joshi, Meghnad, 1977, et al.
(författare)
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Chemokine-Mediated Robust Augmentation of Liver Engraftment: A Novel Approach.
- 2015
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Ingår i: Stem cells translational medicine. - : Oxford University Press (OUP). - 2157-6564 .- 2157-6580. ; 4:1, s. 21-30
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Tidskriftsartikel (refereegranskat)abstract
- Effective repopulation of the liver is essential for successful clinical hepatocyte transplantation. The objective was to improve repopulation of the liver with human hepatocytes using chemokines. We used flow cytometry and immunohistochemistry assays to identify commonly expressed chemokine receptors on human fetal and adult hepatocytes. The migratory capacity of the cells to various chemokines was tested. For in vivo studies, we used a nude mouse model of partial hepatectomy followed by intraparenchymal injections of chemokine ligands at various concentrations. Human fetal liver cells transformed with human telomerase reverse transcriptase were used for intrasplenic cell transplantation. Repopulation and functionality were assessed 4 weeks after transplantation. The receptor CXCR3 was commonly expressed on both fetal and adult hepatocytes. Both cell types migrated efficiently toward corresponding CXC chemokine ligands 9, 10, and 11. In vivo, animals injected with recombinant chemokines showed the highest cell engraftment compared with controls (p < .05). The engrafted cells expressed several human hepatic markers such as cytokeratin 8 and 18 and albumin as well as transferrin, UGT1A1, hepatocyte nuclear factor (1α, 1β, and 4α), cytochrome CYP3A1, CCAAT/enhancer binding protein (α and β), and human albumin compared with controls. No inflammatory cells were detected in the livers at 4 weeks after transplantation. The improved repopulation of transplanted cells is likely a function of the chemokines to mediate cell homing and retention in the injured liver and might be an attractive strategy to augment repopulation of transplanted hepatocytes in vivo.
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| 6. |
- Joshi, Meghnad, 1977, et al.
(författare)
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Fetal liver-derived mesenchymal stromal cells augment engraftment of transplanted hepatocytes.
- 2012
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Ingår i: Cytotherapy. - : Elsevier BV. - 1477-2566 .- 1465-3249. ; 14:6, s. 657-69
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Tidskriftsartikel (refereegranskat)abstract
- One important problem commonly encountered after hepatocyte transplantation is the low numbers of transplanted cells found in the graft. If hepatocyte transplantation is to be a viable therapeutic approach, significant liver parenchyma repopulation is required. Mesenchymal stromal cells (MSC) produce high levels of various growth factors, cytokines and metalloproteinases, and have immunomodulatory effects. We therefore hypothesized that co-transplantation of MSC with human fetal hepatocytes (hFH) could augment in vivo expansion after transplantation. We investigated the ability of human fetal liver MSC (hFLMSC) to augment expansion of phenotypically and functionally well-characterized hFH.
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| 7. |
- Olausson, Michael, 1956, et al.
(författare)
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In vivo application of tissue-engineered veins using autologous peripheral whole blood: A proof of concept study
- 2014
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Ingår i: EBioMedicine. - : Elsevier BV. - 2352-3964. ; 1:1, s. 72-79
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Tidskriftsartikel (refereegranskat)abstract
- Vascular diseases are increasing health problems affecting >25 million individuals in westernized societies. Such patients could benefit fromtransplantation of tissue-engineered vascular grafts using autologous cells. One challenge that has limited this development is the need for cell isolation, and risks associated with ex vivo expanded stem cells. Herewe demonstrate a novel approach to generate transplantable vascular grafts using decellularized allogeneic vascular scaffolds, repopulatedwith peripheralwhole blood (PWB) in vitro in a bioreactor. Circulating, VEGFR-2+/CD45+ and a smaller fraction of VEGFR-2+/CD14+ cells contributed to repopulation of the graft. SEMmicrographs showed flat cells on the luminal surface of the grafts consistentwith endothelial cells. For clinical validation, two autologous PWBtissue-engineered vein conduits were prepared and successfully used for bypass procedures in two pediatric patients. These results provide a proof of principle for the generation of transplantable vascular grafts using a simple autologous blood sample, making it clinically feasible globally.
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| 8. |
- Olausson, Michael, 1956, et al.
(författare)
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Transplantation of an allogeneic vein bioengineered with autologous stem cells: a proof-of-concept study.
- 2012
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Ingår i: Lancet. - 1474-547X. ; 380:9838, s. 230-237
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Tidskriftsartikel (refereegranskat)abstract
- BACKGROUND: Extrahepatic portal vein obstruction can have severe health consequences. Variceal bleeding associated with this disorder causes upper gastrointestinal bleeding, leading to substantial morbidity and mortality. We report the clinical transplantation of a deceased donor iliac vein graft repopulated with recipient autologous stem cells in a patient with extrahepatic portal vein obstruction. METHODS: A 10 year old girl with extrahepatic portal vein obstruction was admitted to the Sahlgrenska University Hospital in Gothenburg, Sweden, for a bypass procedure between the superior mesenteric vein and the intrahepatic left portal vein (meso Rex bypass). A 9 cm segment of allogeneic donor iliac vein was decellularised and subsequently recellularised with endothelial and smooth muscle cells differentiated from stem cells obtained from the bone marrow of the recipient. This graft was used because the patient's umbilical vein was not suitable and other strategies (eg, liver transplantation) require lifelong immunosuppression. FINDINGS: The graft immediately provided the recipient with a functional blood supply (25-30 cm/s in the portal vein and 40 mL/s in the artery was measured intraoperatively and confirmed with ultrasound). The patient had normal laboratory values for 9 months. However, at 1 year the blood flow was low and, on exploration, the shunt was patent but too narrow due to mechanical obstruction of tissue in the mesocolon. Once the tissue causing the compression was removed the graft dilated. We therefore used a second stem-cell populated vein graft to lengthen the previous graft. After this second operation, the portal pressure was reduced from 20 mm Hg to 13 mm Hg and blood flow was 25-40 cm/s in the portal vein. With restored portal circulation the patient has substantially improved physical and mental function and growth. The patient has no anti-endothelial cell antibodies and is receiving no immunosuppressive drugs. INTERPRETATION: An acellularised deceased donor vein graft recellularised with autologous stem cells can be considered for patients in need of vascular vein shunts without the need for immunosuppression. FUNDING: Swedish Government.
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| 9. |
- Patil, Pradeep B, 1982, et al.
(författare)
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CD271 identifies functional human hepatic stellate cells, which localize in pen-sinusoidal and portal areas in liver after partial hepatectomy
- 2014
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Ingår i: Cytotherapy. - : Elsevier BV. - 1465-3249 .- 1477-2566. ; 16:7, s. 990-999
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Tidskriftsartikel (refereegranskat)abstract
- Background aims. Hepatic stellate cells (HSCs) are liver-resident mesenchymal cells involved in essential processes in the liver. However, knowledge concerning these cells in human livers is limited because of the lack of a simple isolation method. Methods. We isolated fetal and adult human liver cells by immunomagnetic beads coated with antibodies to a mesenchymal stromal cell marker (CD271) to enrich a population of HSCs. The cells were characterized by cell cultivation, immunocytochemistry, flow cytometry, reverse-transcription polymerase chain reaction and immunohistochemistry. Cells were injected into nude mice after partial hepatectomy to study in vivo localization of the cells. Results. In vitro, CD271(+) cells were lipid-containing cells expressing several HSC markers: the glial fibrillary acidic protein, desmin, vimentin and alpha-smooth muscle actin but negative for CK8, albumin and hepatocyte antigen. The cells produced several inflammatory cytokines such as interleukin (IL)-6, IL-1A, IL-1B and IL-8 and matrix metalloproteinases MMP-1 and MMP-3 and inhibitors TIMP-1 and TIMP-2. In vivo, fetal CD271(+) cells were found in the pen-sinusoidal space and around portal vessels, whereas adult CD271(+) cells were found mainly in the portal connective tissue and in the walls of the portal vessels, which co-localized with alpha-smooth muscle actin or desmin. CD271(-) cells did not show this pattern of distribution in the liver parenchyma. Conclusions. The described protocol establishes a method for isolation of mesenchymal cell precursors for hepatic stellate cells, portal fibroblasts and vascular smooth muscle cells. These cells provide a novel culture system to study human hepatic fibrogenesis, gene expression and transcription factors controlling HSC regulation.
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| 10. |
- Patil, Pradeep B, 1982
(författare)
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Experimental models with specific approaches to augment human fetal liver cell engraftment
- 2014
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Doktorsavhandling (övrigt vetenskapligt/konstnärligt)abstract
- Experimental models with specific approaches to augment human fetal liver cell engraftment Pradeep Patil Department of Surgery, Institute of Clinical Sciences, Sahlgrenska Academy at University of Gothenburg Background: Liver disease is a common cause of morbidity and mortality worldwide. Orthotopic liver transplantation has so far been the only available therapy for patients with end-stage liver failure. Unfortunately, the availability of donor organs is limited and more than 40% of patients become too sick to survive each year while waiting for liver transplants. Cellular therapy with stem cells and their progeny is a promising new approach to this largely unmet medical need, but is yet to be integrated into the current clinical system. Impediments in cell transplantation are well characterized, but there is lack of reliable solutions, which has limited the use of this technique to act as a bridge (temporary support) to transplantation. Aims: Studies covered under the current thesis are focused on validation and evaluation of reliable cell sources and feasible protocols for enhancing their engraftment and proliferation in animal models. Materials and methods: The mammalian fetal liver contains colony-forming cells with high proliferative potential. The use of human fetal liver cells (hFLCs) is a suitable candidate for the purpose of cell therapy and diagnostics. We have evaluated hFLCs lines as a potential source of stem cells and tested their in vivo functions in a model of liver injury using nude mouse. Results and discussion: This thesis has shown that the regimens of preconditioning (using chemokines) or the co-transplantation (liver cells with mesenchymal stem cells) have the possibility to augment engraftment. Also, manipulating liver cells ex vivo to increase longevity helps in growing cell colonies much faster for many passages to produce a limitless population. It also demonstrates a novel marker to isolate adult or fetal liver stellate cells, which has an important role in immunoregulation and liver fibrosis. Summary: This thesis describes and highlights novel and feasible approaches in liver cell transplantation, with the possibility to improve current clinical protocols. Keywords: cell transplantation, chemokines, SV40, stellate cell, MSCs
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