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 upon the number of copies of donor genome incorporated into the host tissue. We have developed a system for quantitative studies of MT that measures amounts of donor-derived genome in host muscles and estimates the contributions of donor cell survival and proliferation in vivo.


METHODS
[14C]thymidine-labeled, male myoblasts were transplanted into female muscles, providing two donor cell markers, Y chromosome and [14C]. The markers were measured in muscle extracts by slot blotting and scintillation counting, respectively.


RESULTS
In each extract, the amount of Y chromosome was used to quantify donor-derived genome, whereas the radiolabel provided an estimate of cell survival. Furthermore, the different modes of inheritance of the markers meant that proliferation of surviving donor cells was detected as a change in marker ratio.


CONCLUSIONS
This system provides a method for assessing potential improvements of MT.
Beauchamp
J R
JR
Muscle Cell Biology Group, MRC Clinical Sciences Centre, Royal Postgraduate Medical School, Hammersmith Hospital, London, England.
Pagel
C N
CN
Partridge
T A
TA
D000818:Animals; D015415:Biomarkers; D002250:Carbon Radioisotopes; D005260:Female; D018014:Gene Transfer Techniques; D008297:Male; D051379:Mice; D018485:Muscle Fibers, Skeletal / Q000637:transplantation*; D018482:Muscle, Skeletal / Q000378:metabolism; D012588:Scintillation Counting; D012727:Sex Characteristics; D013936:Thymidine / Q000032:analysis; D014998:Y Chromosome
D015415:Biomarkers; D002250:Carbon Radioisotopes; D013936:Thymidine