Isolation of human alveolar macrophages from tumour lung biopsies

Equipment and reagents

• Sterile forceps and scalpel blades

• Sterile gauze

• Sterile heparin!zed saline: 0.85% NaCl, 8.5 U/ml heparin (Sigma Chemical}

Method

1 Immediately place the lung biopsy specimen in sterile heparinized saline at 4 °C.

2 Transfer the tissue to a sterile Petri dish containing 2-4 ml HBSS.

3 Tease and mince the parenchyma using sterile forceps and a scalpel blade.

4 Filter the resulting fragments through several layers of sterile gauze to obtain a single cell suspension.

5 Isolate the mononuclear cells by Ficoll-Hypaque density centrifugation (see Chapter 2).

* Sterile 75 x 15 mm Petri dish

• Hank's balanced salt solution (HBSS) (Gibco BRL)

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