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_recombinant_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 axolotl

with GFP-actin in the

Schwann cells