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What is the Most Likely Candidate for Successful Human Stem Cell Therapy Soonest? • Blood • Muscle • Skin • Why? • How to select a candidate? Are SATELLITE CELLS the same as MYOBLASTS? Stem Cells: Mouse Embryonic Stem Cells http://www.youtube.com/watch?v= 5_9WNwC-4L8&feature=related A muscle cell is a MYOFIBER A myofiber is a SYNCYTIUM What is a SYNCYTIUM? How a muscle works – structure the sarcomere Making a Syncytium Time-lapse Microscopy showing Myoblast Fusion Into Myotubes [Folch lab] http://www.youtube.com/watch?v= cQ-ahxaG8o4 Myoblasts Fusing into a Myotube Beating Human Heart Cells from Embryonic Stem Cells http://www.youtube.com/watch?v= JjVUYChg1M8&feature=related Cycling Cells in the Life of a Muscle Myotube with Satelite Cells Satellite Cells in Muscle Repair Muscle repair and growth – how are they similar and different? Effect of Ageing on Myoblasts Myoblast transfer as a platform technology of gene therapy Peter Law, Tena Goodwin, Qiuwen Fang, George Vastagh, Terry Jordan, Tunja Jackson, Susan Kenny, Vijaya Duggirala, Charles Larkin, Nancy Chase, William Phillips, Glenn Williams, Michael Neel, Tim Krahn, and Randall Holcomb Gene Ther Mol Biol Vol 1, 345-363. March, 1998. Becker dystrophy normal Becker dystrophy Effect of transplanting 50 billion normal myoblasts on enzyme leakage First muscular dystrophy subject ever to walk after wheelchair bound for years. Muscle transplantation between young and old rats: age of host determines recovery • B. M. Carlson and J. A. Faulkner • Am J Physiol Cell Physiol 256: C1262-C1266, 1989 • As compared with age-matched controls, extensor digitorum longus (EDL) muscles autografted in young rats regenerated significantly greater mass (1.8 times) and developed greater maximum contractile force (2.6 times) than EDL muscles autografted in old rats. A cross-age transplantation study showed that the mass and maximum force of old muscles grafted into young hosts were not significantly different from those of young muscles grafted into the same young hosts. Conversely, young muscle grafted into old hosts regenerated no better than old muscles grafted into the same old hosts. We conclude 1) that chronological age alone is not a factor that limits the intrinsic ability of a muscle to regenerate and 2) that the poor regeneration of muscles in old animals is a function of the environment for regeneration provided by the old host. Recombinant DNA Technology http://www.medicalive.net/243_rec ombinant_dna CAUTION!!!!! …some links are fraught with errors http://www.youtube.com/watch?v=YdjvUv-1vCI Green Fluorescent Protein http://gfp.conncoll.edu/ Techniques behind the study • Grafting cells from green axolotl embryos to normal animals before amputation, the researchers could track the GFP to examine the fate of specific cell types in a regenerating limb after amputation in juveniles. • Using these techniques, the researchers looked at four different tissue types: dermis, cartilage, muscle, and Schwann cells - neural tissue that insulates the nerves of the limbs. With the exception of dermal cells, they found that the grafted green cells showed up only in those same tissue types in the regrown limb. Axolotl limb regeneration Regenerated limb labeled with GFP-actin in the Schwann cells axolotl