Initiation of Lymphocyte Culture

1. Two cultures are initiated for each subject.

2. For each culture, add 10 mL of culture medium to two 15-mL centrifuge tubes.

3. Using a Pasteur pipet, inoculate each centrifuge tube with 12 to 15 drops of whole blood. Each centrifuge tube represents a separate lymphocyte culture. Label the tubes.

4. Add 0.15 mL of PHA to each tube and mix well.

5. Incubate the cultures at 37°C for 72 h (in general, the cultures may remain undisturbed for the full 3-d period, although some groups gently agitate the cultures a few times each day).

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