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Biocompatibility and immunocompatibility

IMIC WP 6.1 In vitro immunological properties of polyplexes and polymer membranes WP 6.2 In vivo immunological properties of polyplexes and polymer membranes. Biocompatibility and immunocompatibility. Screening 1. Hemolysis (human red blood cells)

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Biocompatibility and immunocompatibility

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  1. IMICWP 6.1In vitro immunologicalproperties of polyplexes and polymer membranesWP 6.2In vivoimmunological properties of polyplexes and polymermembranes

  2. Biocompatibility and immunocompatibility Screening 1. Hemolysis (human red blood cells) 2. Proliferation (mouse spleen cells, human peripheral blood mononuclear cells – unstimulated and mitogen-stimulated) 3. Nitric oxide production – inflammatory reaction of cells 4. Growth of selected permanent cell lines 10 wt% gelatin type B hydrogels polymerization of methacryamide modified gelatin using e-beam Adherent cell growth – fibroblasts

  3. Fibroblast-type 3T3 mouse cells cultured on gelatin B hydrogel: culture on day 3 (confluent in controls) – tested for viability (metabolic activity) by MTTor XTT assay MTT[3-(4,5-dimethylthiazol-2-yl)-2,5-difenyl tetrazolium bromide ] reduced to insoluble color formazan by mitochondrial dehydrogenases of living cells; solubilization by 10 % SDS, acid propanol, DMSO XTT: reduced form is soluble Alamar blue: suitable for metabolic activity assays, non-toxic – possible to use for continuous monitoring

  4. untreated gelatin

  5. Poly(dimethylaminoethyl-L-glutamine) = PDMAEG (V01) molecular weight: 44000 g/mol mass/charge: 235 g/mol Poly(dimethylaminoethyl-L-glutamine) = PDMAEG (V03) molecular weight: 242000 g/mol mass/charge: 235 g/mol Cationic polymers (UGhent) Biocompatibility tested by hemolysis (human blood) >>> no hemolysis in concentrations 1 mg/ml and lower during 24 and 48 hours

  6. Proliferation of mouse spleen cells: 3 days, 3H-thymidine incorporation (cpm)

  7. LDH (lactate dehydrogenase) assay: non-dividing cells do not produce LDH

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