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Progression of the regenerative process was evident by the occurrence of elongated multinucleated myofibers at days 14 and 30 post transplantation (Figure ?(Figure4).4). The regenerating area gradually extended to occupy 50% ROR1 to 70% of the implants (Figure ?(Figure5)5) Pax7+ cells were detected in all implants, mostly in the periphery. Their number in murine donor tissue was highest at day 7 after implantation (Figure ?(Figure2C).2C). The observed decrease in the number of Pax7+ cells at later time points (day 30) coincided with an increase in myoregeneration in the fresh murine muscle implants (compare Figure ?Figure2C2C with Figure ?Figure55). The regeneration process in the fresh human muscle implants resembled that in the murine implants, albeit at a slower rate (Figures ?(Figures44 and ?and5).5). The extent of regeneration as deduced from the number of myoblasts and regenerated myofibers in the human see more tissue at day 30 and 45 after implantation was lower than in the murine muscle at day 14 post transplantation. Notably, the number of satellite (i.e., Pax7+) cells in the human muscle implants was twice to four times as high as that observed in the murine muscle tissue (Figure ?(Figure2C2C). Regeneration of cryopreserved muscle tissue Because of the logistic problems caused by the irregular supply of human muscle samples and their limited size, we explored the option of using cryopreserved tissue. This approach allows also pooling of samples collected at different dates and from different donors, a strategy that might improve reproducibility. The regeneration of cryopreserved mouse muscle closely resembled that of the fresh implants as judged by HPS and desmin stainings (Figure ?(Figure4).4). The frequency of Pax7+ cells (Figure ?(Figure2C)2C) was not much affected by the freezing procedure, except for day 45 (P www.selleckchem.com/products/MK-2206.html adipose tissue. Degenerating muscle tissue gradually disappeared while the level of newly formed myoblasts/myofibers remained low (Figure ?(Figure4).4). In contrast to the fresh muscle implants, the number of satellite cells decreased with time (Figure ?(Figure2B),2B), suggesting damage to the satellite cell population by the preservation procedure. Contribution of host cells to myoregeneration in subcutaneous muscle implants The skin of a mouse, like that of most rodents, contains a thin skeletal muscle layer named the panniculus carnosus.