Novel tissue culture method: skeletal muscle implantation under gizzard serous membrane of a chick.

A novel method for a long-term culture of skeletal muscle is described. Skeletal muscle pieces from young chicks were implanted under the gizzard serous membrane of the same chicks. Following muscle degeneration, new well-grouped muscle fibers were formed by the fusion of myocytes that differentiated from surviving satellite cells, and the regenerated muscle tissues were maintained in position for longer than 60 days. The implants were in the vital circulatory system, receiving trophic and oxygen supplies, and are completely free from motor nerve innervation and cell contamination with exogenous muscle cells, not as in intra-muscular implantation. Therefore, this tissue culture method should be useful for studying skeletal muscle regeneration and maturation over a long period. Furthermore, osteogenesis and feather development were also found in the implants of embryonic limbs by using the same method. These observations showed that not only skeletal muscle tissues but also other tissues could be cultured under the gizzard serous membrane.