Preparation of microcells by enucleation of micronucleate cells

Microcell-mediated transfer of human DNA repair gene(s) into xeroderma pigmentosum cells.

“…Under these conditions the frequencies of mono-, bi-, tri-and multi-(>_ _ 4) nucleated cells were 81, 8, 4 and 7%, respectively. Although this frequency of micronucleated cells was much less than those reported in rat kangaroo, mouse and rat cells (Ege and Ringertz 1974;Sekiguchi et al 1978), longer treatment or higher concentration of colcemid did not show apparent increase in the frequency. Cytofluometric measurement of Feulgen stained micronuclei revealed that DNA contents of most of micronuclei were smaller than those of intact nuclei and the modal value of the DNA content of micronuclei was about one third of that of control (Fig.…”

“…However, if a part of a normal human chromosome complement is transferred into XP cells, it will be possible to identify the chromosome (s) on which human repair gene (s) is located. Ege and Ringertz (1974) demonstrated that it was possible to prepare microcells by enucleation of micronucleated cells. Gene transfer mediated by the microcells, containing limited number of chromosomes of human or rodent cells, into recipient cells has been reported to be successful (Fournier and Ruddle 1977a;Fournier and Ruddle 1977b;Tourian et al 1978;Sundar Raj et al 1977) .…”

Further Description of a Temperature-Sensitive Mammalian Cell Mutant Exhibiting Micronucleation

“…The formation of micronucleate cells was apparently preceded by an increase in mitotic cell fractions. There was a close resemblance in the time of appearance as well as in morphology between temperature-induced micronuclei in ts 39 cells and colcemid-induced micronuclei in wild-type cells (1,2). In this connection, it is interesting to examine ts 39 cells in relation to the colchicine-sensitivity and colchicine-binding activity of the cell extract.…”

Microcell-mediated transfer of fish chromosomes into mouse cells.

“…The possibility of this method was first demonstrated by Ege and Ringerz (1974) ;Fournier and Ruddle (1977) succeeded in obtaining a microcell hybrid. Subsequently this method has been applied to somatic cell genetics in several ways using mainly mammals.…”

“…The preparation of microcells was made mainly following to the method by Fournier and Ruddle (1977). 10~ cells were placed on two plastic discs cut from a 60 mm tissue culture dish, placed in a 60 mm tissue culture dish, and treated for the micronucleation with 1µM/l of TN-16, 3-(1-anilino-ethylidene)-5-benzylpyrolidine-2,4-dione, for 24 h and then 0.3 µg/ml of colcemid for 48 h. Micronucleated cells on the discs were enucleated by centrifugation at 24000 g *' This work was supported by a grant-in-aid (No .…”