Immunomagnetic purification of rat proximal kidney cells.
A method for producing large quantities of pure proximal tubule cells is described. The procedure involves collagenase perfusion to prepare kidney cells that are separated by centrifugal elutriation to produce a purified cell preparation of proximal tubule cells (PCS). The elutriation-purified cells were treated with a rabbit polyclonal antibody to rat gamma-glutamyl transpeptidase (GGT). The resulting antibody-labelled cells were then incubated with Dynabeads coated with a sheep anti-rabbit IgG antibody. The Dynabeads, which are monodispersed polystyrene beads with a magnetic ferrite core, bind specifically to the antibody-labelled cells. The cell-bead complexes were then harvested with a magnet and washed twice to remove cells not labelled with antibody. The procedure is simple and rapid, and can be used to produce 20-25 x 10(6) cells. The cells exhibit a well defined brush border membrane with increased GGT and alkaline phosphatase enzyme activities. Concomitant with this is a four-fold decrease in hexokinase activity, which demonstrates that contaminating distal and loop of Henle tubule cells have been removed.
Cain
K
K
MRC Toxicology Unit, Woodmansterne Road, Carshalton, Surrey SM5 4EF, UK.
Gurney
J E
JE