Processing of urine and buccal cells

Abstract


The protocol is derived from the WHO/IARC guideline “Common Minimal Technical Standards and Protocols”. The purpose of the document is to provide a framework for the development and coordination of Biological Resource Centers.

Equipment

  • Cryovial
  • Vacutainer , or similar container with a 50-3000 ml capacity

Reagents

  • Liquid Nitrogen , for storage
  • Tris-EDTA buffer

Instructions

    Urine
    1. Plastic or glass containers for collection of urine should be clean and dry, have a 50-3000ml capacity, wide mouth and leak-proof cap, and should be clearly labelled.
    2. When in transit, urine collections should be maintained on ice or refrigerated.
    3. Urine should be aliquoted according to the volume necessary for analysis or storage.
    4. Depending on the analyte to be measured, already during collection or before aliquoting a preservative may be added.
    5. Store urine at -80°C or lower in liquid nitrogen.
    Buccal Cell
    1. A collection kit (containing mouthwash, 50ml plastic tube, plastic biohazard bottle, and courier packaging) is mailed or given to the participant, along with an instruction sheet. The participant is to brush the teeth as usual, rinse the mouth well twice with water, and then wait 2 hours.
    2. They should not eat or drink anything other than water during this time.
    3. After 2 hours, 10ml of commercial mouthwash should be poured into the tube, and then 10ml tap water added. This diluted mouthwash should be placed into the mouth (without swallowing) and swished around vigorously for 30 seconds.
    4. The mouthwash should then be spat back into the plastic tube and the tube should be sealed tightly.
    5. The sample should be sent back to BRC immediately for processing, or stored at 4°C until sent but should be sent within 24 hours.
    6. On arrival at the laboratory, transfer mouthwash to 15ml conical test tubes.
    7. Add 35ml Tris-EDTA to the mouthwash sample and spin at 450g for 5 minutes.
    8. Decant supernatant and discard.
    9. Wash cells twice, each time with 45ml Tris-EDTA.
    10. Resuspend cell pellet in 50μl Tris-EDTA and transfer to 2ml labeled cryovials.
    11. Store sample at -80°C or in liquid nitrogen. Note: the collection of buccal cells can also be conducted with other means such as brushes for example.

    Citation:
    IARC -International agency for research on cancer. Processing of urine and buccal cells. The Molecular Methods database. Wed, 12/19/2012 - 11:27. Acc.nr iyfLt10.