Dopaminergic neuronal conversion from adult rat skeletal muscle-derived stem cells in vitro.
Muscle-derived stem cells reside in the skeletal muscle tissues and are known for their multipotency to differentiate toward the mesodermal lineage. Recent studies have demonstrated their capacity of neuroectodermal differentiation, including neurons and astrocytes. In this study, we investigated the possibility of dopaminergic neuronal conversion from adult rat skeletal muscle-derived stem cells. Using a neurosphere protocol, muscle-derived stem cells form neurosphere-like cell clusters after cultivation as a suspension, displaying an obvious expression of nestin and a remarkable down-regulation of myogenic associated factors desmin, MyoD, Myf5 and myogenin. Subsequently, these neurosphere-like cell clusters were further directed to dopaminergic differentiation through two major induction steps, patterning to midbrain progenitors with sonic hedgehog and fibroblast growth factor 8, followed by the differentiation to dopaminergic neurons with neurotrophic factors (glial cell line-derived neurotrophic factor) and chemicals (ascorbic acid, forskolin). After the differentiation, these cells expressed tyrosine hydroxylase, dopamine transporter, dopamine D1 receptor and synapse-associated protein synapsin I. Several genes, Nurr1, Lmx1b, and En1, which are critically related with the development of dopaminergic neurons, were also significantly up-regulated. The present results indicate that adult skeletal muscle-derived stem cells could provide a promising cell source for autologous transplantation for neurodegenerative diseases in the future, especially the Parkinson's disease.
Key Laboratory for Neurodegenerative Disease of Education Ministry, Department of Physiology and Neurobiology, Capital Medical University, 10# Youanmen, Beijing 100069, People's Republic of China.
Jian
J
Yang
Xuan
X
Wang
Yue
Y
Wang
Zi-Xuan
ZX
Guo
Ding-Zhen
DZ
Luo
Jun
J
Jia
Xiao-Min
XM
Wang
D000818:Animals; D000941:Antigens / Q000032:analysis; D015153:Blotting, Western; D002454:Cell Differentiation / Q000502:physiology; D002478:Cells, Cultured; D003893:Desmin / Q000096:biosynthesis / Q000235:genetics; D059290:Dopaminergic Neurons / Q000378:metabolism / Q000502:physiology*; D005434:Flow Cytometry; D015870:Gene Expression / Q000502:physiology; D007150:Immunohistochemistry; D018485:Muscle Fibers, Skeletal / Q000276:immunology / Q000378:metabolism / Q000502:physiology*; D010300:Parkinson Disease / Q000628:therapy; D012313:RNA / Q000096:biosynthesis / Q000235:genetics; D051381:Rats; D060888:Real-Time Polymerase Chain Reaction; D033581:Stem Cell Transplantation; D013234:Stem Cells / Q000276:immunology / Q000378:metabolism / Q000502:physiology*; D014446:Tyrosine 3-Monooxygenase / Q000378:metabolism
D000941:Antigens; D003893:Desmin; D012313:RNA; D014446:Tyrosine 3-Monooxygenase