Purification of bovine adrenal chromaffin cells by differential plating

A method is described for obtaining highly purified cultures of bovine chromaffin cells from crude adrenomedullary cell suspensions. The method is based on the different adhesiveness of chromaffin and non-chromaffin cells to glass and plastic surfaces (differential plating). Crude suspensions isolated by a modified version of the method described by Livett et al. (1979) (cf. Fenwick et al., 1978) contain 74.4 +/- 7.7% (n = 7) chromaffin cells as determined by electron microscopy. Bringing the cells through 5 steps of differential plating results in cultures that are predominantly composed of chromaffin cells (97.5 +/- 0.85%, n = 8). More than 90% of these cells are viable as judged by trypan blue exclusion and by electron microscopy. Cultures obtained by differential plating contain a significantly lower proportion of non-chromaffin cells than primary cultures both after one week (45.5 +/- 1.2% vs 86.7 +/- 4.6%, n = 3) and after two weeks (85 vs 93%), when grown in Falcon flasks with medium 199 and 20% fetal calf serum, but without mitosis inhibitors. Cultures obtained by the method described in this paper may be profitably employed for studying the contribution of non-chromaffin cells to the functions of chromaffin cells.