Separation of Tumor Cells from Fibroblasts
It is well known that clinical and experimental microbiology made little progress until 1881, when Robert Koch described his method of preparing cultures on solid media. The use of solid media permitted rapid separation of different types of bacteria, and thus allowed the selected organisms to be grown out into pure cultures for further study. The art of tissue culture is somewhat akin to microbiology before the year 1881. It is very difficult to obtain “pure” cultures of selected cell types from most organs and tissues of man and animals. In studies of human tumors, this problem is particularly exasperating; the desired cell is the tumor cell, but the cell usually obtained is the fibroblast from the connective tissue elements associated with the tumor. The technique most widely used to obtain cultures of cells from organs and tissues of mammals utilizes controlled enzyme activity to break down the tissue fragment into a suspension of single cells or small groups of cells. The technique has proved effective for many tissues, particularly those derived from embryos or from organs without much fibrous stroma; however, adult tissues and those with abundant fibrous stroma provide a challenge and a stumbling block for the tissue culturist, who as it were wants the wheat, not the chaff and weeds. The methodology to be described, when tailored to the system being studied, has usually proved effective in separating a high proportion of tumor cells from the “contaminating” fibroblasts.
KeywordsMinimal Essential Medium Uninfected Cell Interface Zone Buoyant Density Ficoll Gradient
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