Isolation of neuroglia and inflammatory cells from neural tissue

In neural tissue there are numerous types of “support cells” which are grouped under the term “Neuroglia” and this Application Sheet is concerned with just two of them. Iodixanol gradients have been used to purify neuroglial cells and also to remove myelin from disaggregated CNS tissue prior to the use of antibody-bound beads to purify oligodendrocytes. 

Microglial cells have been isolated both from mouse brain, using the standard methodology of tissue mincing followed by papain [1] or trypsin digestion [2] to disaggregate the cells. They have also been isolated from culture after a six-stage in vitro differentiation of mouse embryonic stem cells [2].

 The latter involves expansion of undifferentiated stem cells, followed by generation of embryoid bodies, selection and expansion of nestin-positive cells, differentiation into neurons and expansion of microglia. Details of this methodology are beyond the scope of this Application Sheet and the reader is referred to ref 2 for details.