Application of flow cytometry to analyses of cultured XXXX @entity11 and fetal brain cells.

@entity1 @entity538 @entity11 @entity816 @entity1604 @entity463 @entity1098 @entity3271
Methods are described to study cell surface and cytoplasmic antigens of cultured @entity1 @entity11 , fetal brain cells and fibroblasts using flow cytometry. This required harvesting the cultured cells with @entity1098 or mild trypsin treatment and fixation in 4% @entity1604 prior to staining for @entity538 ( @entity538 ) and fibronectin using indirect immunofluorescence. At passage 10, 38% of fetal brain cells [CHII] were @entity538 -positive but at passage 14 only 3.5% expressed @entity538 . Two @entity463 and an @entity816 had 38.8%, 6.7% and 81.3% @entity538 -positive cells, respectively. Of the 10(4) cells studied, 91.6%, 79.1% and 40.8% were fibronectin-positive for @entity463 multiforme [12-18], @entity3271 [12-10] and fetal brain [CHII] cells, respectively. Two fibroblast lines had 33.5% and 43.1% of the cells expressing fibronectin. The validity of these results was confirmed by staining for @entity538 and fibronectin using peroxidase-antiperoxidase and immunofluorescence microscopy. Using low angle forward light scatter to estimate cell size and gating techniques it was found that @entity538 -positive CHII and @entity816 cells were generally larger than @entity538 -negative cells of the same type. No correlation between cell size and fibronectin expression was found for @entity463 [12-18] cells. These results demonstrate the validity of the described methods and illustrate some specific applications and the potential value of flow cytometry to neurooncology.
