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									University of Sheffield Department of Engineering Materials

8 December2005

CoSHH Assessment – Experimental
CoSHH Ref No: Describe Procedure:
Isolation of Stromal Cells and Urothelial Cells from human Prostate and Bladder. Human Urothelium is collected from theatres as Prostate or Bladder. The Urothelium is then cut into small pieces before incubation in the chosen enzyme allowing the cells to be isolated.

Lab where procedure will be carried out: 2nd floor laboratory, Kroto Research Institute

List ALL Substances and Quantities product):

Hazards Identified:Used (including

EDTA (0.02% solution) (3ml) May cause skin and eye irritation. Maybe harmful if swallowed or inhaled. Keratinocyte Serum Free Media (K-SFM) May cause eye and skin irritation. May be harmful if inhaled or swallowed. May be (500ml) harmful if absorbed through the skin. Human recombinant Epidermal Growth May be irritating to eyes, respiratory system and skin. Factor (EGF) (5ng/ml) Bovine pituitary extract (BPE) (50μg/ml) May cause eye and skin irritation. May be harmful if inhaled or swallowed. May be harmful if absorbed through the skin. Hepes Buffer Solution (10mls) May cause eye and skin irritation. Mildly irritating to mouth, throat and stomach. Can cause abdominal discomfort. May cause skin and eye irritation. May cause nausea or vomiting. HBSS (30mls) Due to low % of toxic material (<0.1%), this material is of low risk. May be harmful if Dulbecco’s Modified Eagles Medium ingested. (DMEM) (500ml) May be harmful if ingested. Foetal Bovine Serum (50mls) Harmful Collagenase Biohazard – may contain known infectious hazards. Human Urothelium, blood and cells May be harmful if inhaled, ingested or in contact with skin Penicillin (5mls) May be harmful if inhaled, ingested or in contact with skin. Fungizone (2.5 mls) Biohazard. May cause human disease. Harmful in contact with skin and if swallowed. Cholera Toxin (0.5mls) Irritating to eyes, respiratory system and skin. Contact with acids liberates a very toxic gas. May cause sensitization by inhalation and skin contact. Aprotinin

See to be 02/069/JWH Particular Control/safety MeasuresCoSHHadopted: (include where and how substances are stored) Possible source eyes of all substances Avoid ingestion, inhalation or contact with skin and of infectious diseases i.e. HIV, Hep B Wear gloves, lab coat, safety spectacles when handling material Further cell Carry out procedure in Class II Safety Cabinet culture covered in CoSHH 00/032/RMF 01/018/JWH Substances to be stored in fridge/freezer as appropriate with lids tightly closed

Emergency Procedures:

a) Spillage Wipe any spillages with 70% alcohol and dispose in yellow
biohazard bins.

b) Fire

All substances are non-flammable. Use extinguisher appropriate for surroundings. EDTA emits toxic fumes under fire conditions.

Information Sources:
Sigma-Aldrich, Invitrogen and Biowest Ltd (formally Labtech International) Materials and Safety Data Sheets (MSDS).

Could a Less Hazardous Substance be used? If so, why not use it? No

First Aid Treatment:
All workers should be immunised for Hep B Inhalation – Remove to fresh air. If breathing becomes difficult seek medical advice. Exposure to eye – Flush with eye wash for at least 15 minutes and seek medical advice. Exposure to skin – Wash with water for 15 minutes, if irritation develops seek medical advice. Ingestion – Seek medical advice stating exact substances that you have been exposed to.

Waste Disposal Procedures:
Sharps in sharp bin Human Urothelium – special compartment in -20 freezer ready for incineration Inactivate solution (denaturing any active components, such as proteins) by placing in bleach (bleach tablet in water, at least 2500ppm chlorine). Flush waste down the sink with lots of running water (to avoid corrosion of steel sink).

Name of Assessor: Maggie Glover Status of Assessor: Research Technician

I have read this document and approve the above risk assessment


Signed:________________________________ Date:____06/12/05____________________________ Date:_____________________________________ __

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