Mk at210d

C57BL/6 pregnant mice at 14.5 dpc and 16.5 dpc were anaesthetized using 1.0% isoflurane and an animal anesthetizer device (MK-AT210D, Muromachi Kikai Co., LTD, Tokyo, Japan). Mice were then placed on the plate warmed to 37°C, their abdominal area was shaved, and the skin and abdominal wall were incised. The uterine wall was then cut on the side opposite the placenta. Glass needles were prepared from glass capillary tubes (Narishige, Tokyo, Japan) using a micropipette puller (PN-30, Narishige). Under a microscope, 3 μl of a cell suspension containing 1.4×10⁵ FL EGFP(+) CD45(+) c-Kit(+) cells obtained at 14.5 dpc or 5×10³ muscle tissue EGFP(+) CD45(+) c-Kit(+) cells obtained at 16.5 dpc (both from EGFP Tg mouse embryos) were transplanted into corresponding tissues of C57BL/6 recipient embryos. In some experiments, FL CD45(+) c-Kit(+) cells obtained from C57BL/6 mouse embryos at 14.5 dpc were labeled using a Qtracker^® 585 Cell Labeling Kit (Ambion), according to the manufacturer’s protocol, and 1.4×10⁵ cells were transplanted into corresponding tissues of C57BL/6 recipient embryos. The abdominal wall and skin were then sutured using 6/0 silk (Teleflex, Philadelphia, PA). After 24 or 72 hours, embryos were collected and analyzed for EGFP expression or Qdot585 fluorescence in muscle tissue and BM by flow cytometry and/or immunohistochemistry.