Abstract
This chapter describes the methods for dipteran cell cultures that lead to the establishment of cell lines which are predominantly diploid. The chapter discusses the development of the primary cultures and the establishment of diploid lines. The trypsin serves only to loosen the matrix among the cells as visually there is no difference in the appearance of the fragments before and after treatment. The explants behave in one of two ways: either individual cells or groups of cells drop to the floor of the flask and form colonies or growth takes place in the form of hollow, cellular spheres or vesicles issuing from the cut ends of the embryonic or larval fragments. If the former condition prevails, the colonies are allowed to grow until they reach a few millimeters in diameter. The medium is then partially renewed and the colonies pipetted off the bottom of the flask. If growth is in the form of cellular vesicles, they may dislodge from the fragments by themselves or must be excised with tungsten needles. In the latter case, both fragments and vesicles are returned to the original flask to increase the cell density.
Published Version
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