Macrophage subpopulations located in different tissues are interesting to study since they are adapted to specialized activity within their local environment. Because they are closely associated with surrounding tissues, mechanical and/or enzymatic digestion is often necessary. Macrophages can be recovered from liver, gut, brain, bone, and spleen with varying degrees of time, purity, viability, and yield. In other tissues, macrophage experimentation is mostly limited to histological techniques.
To make single cell suspension of splenocytes, only gentle disruption is required and fairly large numbers of macrophages can be obtained from a single mouse spleen and isolated by Percoll gradient centrifugation (30, 31), as described in Protocol 10. Isolation of human splenic macrophages requires a more rigorous protocol of tissue dissociation and is discussed in Protocol 11.
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