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Isolation of fibroblast derived extracellular vesicles using magnetic beads and click chemistry

Scholten, J.N. (2023) Isolation of fibroblast derived extracellular vesicles using magnetic beads and click chemistry.

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Abstract:Hepatocellular carcinoma (HCC) is one of the most common types of cancer worldwide. Early diagnosis leads to higher survival rates. However, a sensitive method to detect HCC in early stages is not yet available. HCC often develops via liver fibrosis. Activated fibroblasts play a role in this process. Extracellular vesicles (EVs) are released from cells and can provide useful information about disease progression. Immunoaffinity-based techniques, such as using magnetic beads and click chemistry with an antibody, provide the isolation of specific types of EVs. The aim of this research was to isolate fibroblast derived EVs using magnetic beads and click chemistry, to detect liver fibrosis in early stages. To achieve this, fibroblast specific surface markers were identified by cell stainings and Dot blot. EVs were characterised by Calcein stainings and Dynamic Light Scattering. Finally, the conjugation with magnetic beads was performed. For the LX-2 cells used here, no fibroblast specific surface marker was identified. Human primary prostate fibroblasts (hPrFs) were found to express Fibroblast activation protein-α (FAP). Conjugation for hPrF EVs with FAP antibody showed the ability of the system to isolate fibroblast derived EVs using magnetic beads and click chemistry. However, as efficiency was low, further research is needed.
Item Type:Essay (Bachelor)
Faculty:TNW: Science and Technology
Subject:35 chemistry, 42 biology, 44 medicine
Programme:Biomedical Technology BSc (56226)
Link to this item:https://purl.utwente.nl/essays/96331
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