The development of three cell lines initiated from the late embryonic stages of Drosophila melanogaster is described. The primary cultures consisted of trypsinized fragments from embryos 20–24 h old. The length of time between primary culture and subsequent subculture varied from 8 months for the first line to 3 weeks for the third. All three lines have been maintained in vitro for more than a year. The characteristics of each line are given and evidence is presented that at least one line is derived from imaginal disc cells. A few comments on insect tissue culture in general are also made.

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