Lecture 5 Safety, Bioethics Validation

Lecture 5 Safety, Bioethics Validation

  • SOP
  • Safety
    • Biosafety levels
      • 1 well characterized agents not known to cause ds
      • 2 moderate-risk agents moderate danger to personnel & environment
      • 3 indigenous/exotic potential respiratory transmission & serious/lethal infection
      • 4 dangerous exotic agents high risk of life threatening ds, aerosol transmission, unknown risk of transmission, most stringent
    • MSDS
    • General safety
      • trained operator
      • supervisor ensure proper procedure & attire
      • proper disposal of glassware & sharps for incineration
    • Chemical toxicity
      • use liquid detergent, avoid powdered
      • disinfectant hypochlorite bleach skin
      • dimethyl sulphoxide penetrate skin
      • mutagen, carcinogen, cytotoxic drugs in fume hood/class 2
      • OSH officer had detailed handling method
    • CO2, O2, N2
      • CO2, N2 cause asphyxiation
      • O2 cause fire
      • evacuation route
      • N2 -196C→frostbite, asphyxiation, explosion
      • use gloves when handling
    • Fire & burns
      • keep alcohol in narrow/plastic bottles, minimize volume
      • do not return burnt sterilized instruments to alcohol when still alight, always keep a damp cloth nearby to smother flames
      • source of risk
        • high temp equipment
        • handling items removed from high temp equipment
        • naked flames (Bunsen burner)
    • Ionizing radiation
      • ingestion→low energy beta emitters (3H soluble); class 2 BSC
      • irradiation from labeled reagents→beta & gamma emitters (32P, 125I, 131I, 51Cr); use lead/perspex
      • irradiation from high energy sources→x ray (60Co) UV; consult radiology
  • Biohazards
    • high grade pathogens (anthrax spores) - fatal
  • Biopsies, genetic manipulation, disposal & fumigation
    • obtain consent from donors
    • transport specimens in double wrapped containers, log all samples, use secure refrigeration marked with a biohazard label
    • carry out dissection in separate class 2 BSC
    • transformation follow SOP as unknown infectivity
    • disposal involve autoclave & hypochlorite
    • fumigation with formaldehyde/H peroxide
  • Bioethics
    • animal & human tissue-legislation
    • ethics committees
    • animal welfare consent forms must be signed
    • confidentiality & acknowledgement-legal proceedings
    • informed consent
    • stem cell research ethics
    • genetic modification→CRISPR
    • confidentiality & data protection
    • commercialization & intellectual property
  • Validation
    • cell line authentication
      • confirm identity
      • prevent cross-contamination
    • mycoplasma testing
    • fx assays→ensure consistency across passages
    • QC measures
    • GLP & GMP
    • validation of culture media & reagents→lot-lot variation monitor
  • Validation of cell lines
    • DNA fingerprint/profiling
    • provenance involve proper recording of H/O cell line
    • test contamination→culture w/o Ab, Hoescht stain for mycoplasma, PCR for virus
    • cell bank→ATCC, EAECC, Riken
    • new user always use new validated stock of cells

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