A DUAL-MARKER SYSTEM FOR QUANTITATIVE STUDIES OF MYOBLAST TRANSPLANTATION IN THE MOUSE

Background. Myoblast transplantation (MT) is a potential approach for gene transfer into skeletal muscle, the efficiency of which depends up on the number of copies of donor genome incorporated into the host tis sue, We have developed a system for quantitative studies of MT that me asures amounts of donor-derived genome in host muscles and estimates t he contributions of donor cell survival and proliferation in vivo. Met hods. [C-14]thymidine-labeled, male myoblasts were transplanted into f emale muscles, providing two donor cell markers, Y chromosome and [C-1 4], Th, markers were measured in muscle extracts by slot blotting and scintillation counting, respectively. Results. In each extract, the am ount of Y chromosome was used to quantify donor-derived genome, wherea s the radiolabel provided an estimate of cell survival, Furthermore, t he different modes of inheritance of the markers meant that proliferat ion of surviving donor cells was detected as a change in marker ratio. Conclusions. This system provides a method for assessing potential imp rovements of MT.