A method for obtaining suspensions of living cells from the fixed tissues,and for the plating out of individual cells

Abstract

The method depends on the ability of living tissue cells to withstand tryptic digestion. It is applicable to such cells as will proliferate in vitro. Bits of tissue are cultivated in plasma, according to Burrows's modification of Harrison's technic, and when growth is well under way the preparation is flooded with trypsin dissolved in Locke's solution. Under the influence of this fluid the growing cells contract into spheres, and with the digestion of the fibrin network they are liberated. Suspensions of individual cells are thus obtained comparable to suspensions of leukocytes. When washed and plated anew in plasma the cells put forth processes and proliferate. The digestion and plating can be repeated. The method is most successful with tissues that grow loosely in strands or a network,—sarcoma, choroid, endothelium (?), connective tissue,—in distinction from those proliferating in sheets, as do the epithelial tissues. The cells of these latter are usually liberated in clumps, not as individuals.