Neurotrophin-directed differentiation of human adult marrow stromal cells to dopaminergic-like neurons.
Marrow-isolated adult multilineage inducible (MIAMI) cells were differentiated in vitro to neuronal cells in a neurotrophin-dependent fashion. After induction, the cells revealed electrophysiological features similar to those observed in mature neurons. Primary early passage human MIAMI cells without any type of co-cultures with other cell types were used. The developmental program involved a multi-step process requiring the concerted action of brain-derived neurotrophic factor, nerve growth factor and depended on neurotrophin-3, after basic fibroblast growth factor withdrawal. MIAMI-derived neuron-like cells sequentially expressed the neuronal markers, developed a complex neurite outgrowth and arborization, and acquired electrophysiological characteristics similar to those observed in mature neurons. The young and old MIAMI-derived neuronal cells developed both inward and outward currents upon depolarization, similar to those observed in normal neurons. These results represent the earliest evidence that neurotrophin-3 can direct the differentiation of non-neural stem cells from human adult bone marrow stroma to neuron-like cells in vitro. Supplementing the aforementioned multi-step process with sonic hedgehog, fibroblast growth factor 8, and retinoic acid increased the expression of molecules involved in dopaminergic differentiation and of tyrosine hydroxylase, the rate limiting enzyme of dopamine synthesis. MIAMI cells from young and old individuals represent autologous human cell populations for the treatment of disorders of the skeletal and nervous systems and for applications in cell therapy and reparative medicine approaches.
Tatard
Valerie M
VM
INSERM Unit 646, Ingénierie de la Vectorisation Particulaire, 10, rue André Bocquel, 49100 Angers, Centre Hospitalier Universitaire d'Angers, 4 rue Larrey, France.
D'Ippolito
Gianluca
G
Diabira
Sylma
S
Valeyev
Alexander
A
Hackman
John
J
McCarthy
Micheline
M
Bouckenooghe
Thomas
T
Menei
Philippe
P
Montero-Menei
Claudia N
CN
Schiller
Paul C
PC
D000293:Adolescent; D000328:Adult; D053687:Adult Stem Cells / Q000166:cytology / Q000502:physiology; D000367:Age Factors; D000368:Aged; D001854:Bone Marrow Cells / Q000166:cytology / Q000502:physiology*; D002454:Cell Differentiation; D002478:Cells, Cultured; D002648:Child; D002675:Child, Preschool; D004298:Dopamine / Q000378:metabolism*; D005260:Female; D051524:Fibroblast Growth Factor 8 / Q000494:pharmacology; D053823:Hedgehog Proteins / Q000494:pharmacology; D006801:Humans; D008297:Male; D008875:Middle Aged; D016501:Neurites / Q000502:physiology; D009474:Neurons / Q000166:cytology / Q000378:metabolism / Q000502:physiology*; D020933:Neurotrophin 3 / Q000494:pharmacology / Q000502:physiology*; D011994:Recombinant Proteins / Q000494:pharmacology; D017154:Stromal Cells / Q000166:cytology / Q000502:physiology*; D014212:Tretinoin / Q000494:pharmacology; D014446:Tyrosine 3-Monooxygenase / Q000378:metabolism
C496456:FGF8 protein, human; D053823:Hedgehog Proteins; D020933:Neurotrophin 3; D011994:Recombinant Proteins; D051524:Fibroblast Growth Factor 8; D014212:Tretinoin; D014446:Tyrosine 3-Monooxygenase; D004298:Dopamine