Generating neuron-like cells from BM-derived mesenchymal stromal cells in vitro.
BACKGROUND
The multipotency of stromal cells has been studied extensively. It has been reported that mesenchymal stromal cells (MSC) are capable of differentiating into cells of multilineage. Different methods and reagents have been used to induce the differentiation of MSC. We investigated the efficacy of different growth factors in inducing MSC differentiation into neurons.


METHODS
MSC from human BM were isolated and cultured in media supplemented with 10% FBS. These cells were identified and later induced to differentiate into neuron-like cells using different neurotrophic factors. Three different growth factors were used, either alone or in combination: brain-derived neurotrophic factor, epidermal growth factor and neural growth factor.


RESULTS
After 10 days of culture, MSC showed neuron-like morphologic changes. Immunostaining showed that these cells expressed markers for neurons (growth-associated protein-43, neuron-specific nuclear protein and neurofilament 200 kDa) and expression of these markers suggested the transition of immature stages to more mature stages of neuron-like cells.


CONCLUSIONS
Our results show that BM-derived MSC can differentiate not only into target cells of mesodermal origin but also neuron-like cells of ectodermal origin. The findings show that a combination of growth factors is more effective in inducing MSC into neuron-like cells.
Choong
P-F
PF
Department of Pathology, National University of Malaysia, Kuala Lumpur, Malaysia. erychoong@yahoo.com
Mok
P-L
PL
Cheong
S-K
SK
Leong
C-F
CF
Then
K-Y
KY
D017667:Adipocytes / Q000166:cytology / Q000378:metabolism; D000469:Alkaline Phosphatase / Q000032:analysis / Q000235:genetics; D034961:Antigens, Nuclear / Q000032:analysis / Q000235:genetics; D001854:Bone Marrow Cells / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D019208:Brain-Derived Neurotrophic Factor / Q000494:pharmacology; D018929:Cell Culture Techniques / Q000379:methods; D002454:Cell Differentiation / Q000187:drug effects; D002478:Cells, Cultured; D019902:Chondrocytes / Q000166:cytology / Q000378:metabolism; D004815:Epidermal Growth Factor / Q000494:pharmacology; D005434:Flow Cytometry; D019922:GAP-43 Protein / Q000032:analysis / Q000235:genetics; D006801:Humans; D007150:Immunohistochemistry; D016130:Immunophenotyping; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D009414:Nerve Growth Factors / Q000494:pharmacology; D009419:Nerve Tissue Proteins / Q000032:analysis / Q000235:genetics; D016900:Neurofilament Proteins / Q000032:analysis / Q000235:genetics; D009474:Neurons / Q000166:cytology* / Q000378:metabolism; D010006:Osteoblasts / Q000166:cytology / Q000378:metabolism; D015675:Osteocalcin / Q000032:analysis / Q000235:genetics; D053495:Osteopontin / Q000032:analysis / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction
D034961:Antigens, Nuclear; D019208:Brain-Derived Neurotrophic Factor; D019922:GAP-43 Protein; D009414:Nerve Growth Factors; D009419:Nerve Tissue Proteins; D016900:Neurofilament Proteins; C506808:neuronal nuclear antigen NeuN, human; D015675:Osteocalcin; D053495:Osteopontin; C051774:neurofilament protein H; D004815:Epidermal Growth Factor; D000469:Alkaline Phosphatase