| 1. |
- Larsson, Johanna, et al.
(author)
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Säkerhetskultur och självkörande fordon och maskiner
- 2024
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Reports (other academic/artistic)abstract
- Självkörande fordon inom transportsektorn befinner sig ännu i utvecklingsstadiet. Att införa självkörande fordon och maskiner i befintliga verksamheter innebär ofta förändringar både i organisationen och den fysiska miljön och kan även innebära nya risker. I detta sammanhang kan säkerhetskulturen, både hos utvecklare och hos användare, spela viktig roll för att självkörande fordon och maskiner ska fungera säkert och effektivt i olika verksamheter. Projektets mål har varit att utveckla metoder för att förbättra säkerhetskultur där människor och automatiserad teknik samverkar som agenter i ett gemensamt system, samt att utveckla mätverktyg där hållbarhet, jämställdhet och säkerhet utvärderas för införande av självkörande fordon och maskiner. Projektet har utgått ifrån fallstudier från två olika domäner – självkörande bussar och självkörande industritruckar. Intervjuer har genomförts med utvecklare, kunder och slutanvändare. En enkät har tagits fram att mäta säkerhetskultur, jämställdhetskultur och hållbarhetskultur i organisationer som utvecklar självkörande fordon. Utöver detta har data från incidentrapporter analyserats. Lärdomarna från resultaten och projektdeltagarnas tidigare erfarenheter har resulterat i ett första utkast av en processmodell där säkerhetskultur integreras i utvecklingen av självkörande fordon och maskiner. Intervjuerna med utvecklare och kunder av självkörande fordon visade att säkerhetskultur inte var ett etablerat begrepp vare sig hos utvecklarna eller hos kunderna och att det därför inte var en faktor som man medvetet beaktade. Lärdomar från enkäten var att det finns skillnader mellan produktföretag och som markant påverkar formuleringar av frågeställningar. Det gick inte att fastställa om det föreligger kopplingar mellan hållbarhets-, jämställdhets- och säkerhetskultur. Fallstudien med bussarna visade bland annat att kund och leverantör pratar om olika typer av säkerhet samt att säkerheten ofta, men inte alltid, är prioriterad över effektiviteten. Utifrån analyserna av incidentdata från självkörande bussar samt förarlösa industritruckar har en lista med förslag på nyckeltal för att kunna analysera incidenter med självkörande fordon tagits fram inom projektet.
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| 2. |
- Brolen, Gabriella, et al.
(author)
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Hepatocyte-like cells derived from human embryonic stem cells specifically via definitive endoderm and a progenitor stage
- 2010
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In: Journal of Biotechnology. - : Elsevier BV. - 1873-4863 .- 0168-1656. ; 145:3, s. 284-294
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Journal article (peer-reviewed)abstract
- Human embryonic stem cells offer a potential unlimited supply for functional hepatocytes, since they can differentiate into hepatocyte-like cells displaying a characteristic hepatic morphology and expressing various hepatic markers. These cells could be used in various applications such as studies of drug metabolism and hepatotoxicity, which however, would require a significant expression of drug metabolizing enzymes. To derive these cells we use a stepwise differentiation protocol where growth- and maturation factors are added. The first phase involves the formation of definitive endoderm. Next, these cells are treated with factors known to promote the induction and proliferation towards hepatic progenitor cell types. In the last phase the cells are terminally differentiated and maturated into functional hepatocyte-like cells. The cultures were characterized by analysis of endodermal or hepatic markers and compared to cultures derived without induction via definitive endoderm. Hepatic functions such as urea secretion, glycogen storage, indocyanine green uptake and secretion, and cytochrome P450-expression and activity were evaluated. The DE-Hep showed a hepatocyte morphology with sub-organized cells and exhibited many liver-functions including transporter activity and capacity to metabolize drugs specific for important cytochrome P450 sub-families. This represents an importantstep in differentiation of hESC into functional hepatocytes. (C) 2009 Elsevier B.V. All rights reserved.
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| 3. |
- Lundqvist, Magnus, et al.
(author)
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Solid-phase cloning for high-throughput assembly of single and multiple DNA parts
- 2015
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In: Nucleic Acids Research. - : Oxford University Press (OUP). - 0305-1048 .- 1362-4962. ; 43:7
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Journal article (peer-reviewed)abstract
- We describe solid-phase cloning (SPC) for high-throughput assembly of expression plasmids. Our method allows PCR products to be put directly into a liquid handler for capture and purification using paramagnetic streptavidin beads and conversion into constructs by subsequent cloning reactions. We present a robust automated protocol for restriction enzyme based SPC and its performance for the cloning of >60 000 unique human gene fragments into expression vectors. In addition, we report on SPC-based single-strand assembly for applications where exact control of the sequence between fragments is needed or where multiple inserts are to be assembled. In this approach, the solid support allows for head-to-tail assembly of DNA fragments based on hybridization and polymerase fill-in. The usefulness of head-to-tail SPC was demonstrated by assembly of >150 constructs with up to four DNA parts at an average success rate above 80%. We report on several applications for SPC and we suggest it to be particularly suitable for high-throughput efforts using laboratory workstations.
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| 4. |
- Sivertsson, Louise, et al.
(author)
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Hepatic differentiation and maturation of human embryonic stem cells cultured in a perfused three-dimensional bioreactor
- 2013
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In: Stem Cells and Development. - : Mary Ann Liebert. - 1547-3287 .- 1557-8534. ; 22:4, s. 581-594
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Journal article (peer-reviewed)abstract
- Drug-induced liver injury is a serious and frequently occurring adverse drug reaction in the clinics and is hard to predict during preclinical studies. Today, primary hepatocytes are the most frequently used cell model for drug discovery and prediction of toxicity. However, their use is marred by high donor variability regarding drug metabolism and toxicity, and instable expression levels of liver-specific genes such as cytochromes P450. An in vitro model system based on human embryonic stem cells (hESC), with their unique properties of pluripotency and self-renewal, has potential to provide a stable and unlimited supply of human hepatocytes. Much effort has been made to direct hESC toward the hepatic lineage, mostly using 2-dimensional (2D) cultures. Although the results are encouraging, these cells lack important functionality. Here, we investigate if hepatic differentiation of hESC can be improved by using a 3-dimensional (3D) bioreactor system. Human ESCs were differentiated toward the hepatic lineage using the same cells in either the 3D or 2D system. A global transcriptional analysis identified important differences between the 2 differentiation regimes, and we identified 10 pathways, highly related to liver functions, which were significantly upregulated in cells differentiated in the bioreactor compared to 2D control cultures. The enhanced hepatic differentiation observed in the bioreactor system was also supported by immunocytochemistry. Taken together, our results suggest that hepatic differentiation of hESC is improved when using this 3D bioreactor technology as compared to 2D culture systems.
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| 5. |
- Tegel, Hanna, et al.
(author)
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High throughput generation of a resource of the human secretome in mammalian cells
- 2020
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In: New Biotechnology. - : Elsevier BV. - 1871-6784 .- 1876-4347. ; 58, s. 45-54
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Journal article (peer-reviewed)abstract
- The proteins secreted by human tissues and blood cells, the secretome, are important both for the basic understanding of human biology and for identification of potential targets for future diagnosis and therapy. Here, a high-throughput mammalian cell factory is presented that was established to create a resource of recombinant full-length proteins covering the majority of those annotated as 'secreted' in humans. The full-length DNA sequences of each of the predicted secreted proteins were generated by gene synthesis, the constructs were transfected into Chinese hamster ovary (CHO) cells and the recombinant proteins were produced, purified and analyzed. Almost 1,300 proteins were successfully generated and proteins predicted to be secreted into the blood were produced with a success rate of 65%, while the success rates for the other categories of secreted proteins were somewhat lower giving an overall one-pass success rate of ca. 58%. The proteins were used to generate targeted proteomics assays and several of the proteins were shown to be active in a phenotypic assay involving pancreatic beta-cell dedifferentiation. Many of the proteins that failed during production in CHO cells could be rescued in human embryonic kidney (HEK 293) cells suggesting that a cell factory of human origin can be an attractive alternative for production in mammalian cells. In conclusion, a high-throughput protein production and purification system has been successfully established to create a unique resource of the human secretome.
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| 6. |
- Uhlén, Mathias, et al.
(author)
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The human secretome
- 2019
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In: Science Signaling. - : American Association for the Advancement of Science (AAAS). - 1945-0877 .- 1937-9145. ; 12:609
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Journal article (peer-reviewed)abstract
- The proteins secreted by human cells (collectively referred to as the secretome) are important not only for the basic understanding of human biology but also for the identification of potential targets for future diagnostics and therapies. Here, we present a comprehensive analysis of proteins predicted to be secreted in human cells, which provides information about their final localization in the human body, including the proteins actively secreted to peripheral blood. The analysis suggests that a large number of the proteins of the secretome are not secreted out of the cell, but instead are retained intracellularly, whereas another large group of proteins were identified that are predicted to be retained locally at the tissue of expression and not secreted into the blood. Proteins detected in the human blood by mass spectrometry-based proteomics and antibody-based immuno-assays are also presented with estimates of their concentrations in the blood. The results are presented in an updated version 19 of the Human Protein Atlas in which each gene encoding a secretome protein is annotated to provide an open-access knowledge resource of the human secretome, including body-wide expression data, spatial localization data down to the single-cell and subcellular levels, and data about the presence of proteins that are detectable in the blood.
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