Preparation of Metabolically Active Cell Suspensions From Wool Roots

Abstract

A method is described for the preparation of metabolically active cell suspensions from plucked wool roots using the proteolytic enzyme trypsin. The suspensions consist of a heterogeneous population of cells which appear similar in morphology to follicle bulb cells, differentiating keratinocytes and possibly cells of the inner root sheath. The concentrations of trypsin and of inorganic ions for optimum activity of the suspensions have been determined, and the inclusion of EGTA was found to increase the yield of cells. The cell suspensions incorporate [14C]leucine, [3H]uridine and [3H]thymidine into acid-insoluble products, and are sensitive to the action of cycloheximide and emetine, but not to chloramphenicol or rifampin. Autoradiography has shown that the cells believed to be derived from the follicle bulbs show the greatest activity.