Flow cytometric analysis of granulosa cells from follicular fluid after follicular stimulation.

Granulosa cells are not easily accessible, unless they are examined in follicular fluid after oocyte retrieval. These samples are usually contaminated with blood. We have set up a general technique for analysis of granulosa cells without physically separating them from blood cells. The sample is stained with CD45, which is a pan-leukocyte marker, and granulosa cells are consecutively selected as CD45 negative during flow cytometric analysis. Analysis of forward scatter of the granulosa cells, which is correlated to cell size, shows a wide size range throughout the whole population rather than two distinct populations as previously suggested.