Differentiation of mesenchymal stem cells into dopaminergic neuron-like cells in vitro.
OBJECTIVE
To explore the way to induce mesenchymal stem cells (MSCs) to differentiate into dopaminergic neurons in vitro.


METHODS
MSCs were obtained from rat bone marrow, cultured and passaged. MSCs used in this experiment had multipotency, which was indirectly proved by being induced to differentiate into chondrocytes and adipocytes. MSCs were cultured in medium containing 0.5 mmol/L IBMX for 2 days. Then the medium was replaced with induction medium, which contained GDNF, IL-1beta, mesencephalic glial-cell-conditioned medium and flash-frozen mesencephalic membrane fragments. The surface markers of the differentiated neurons, such as NSE, nestin, MAP-2a, b and TH were detected by immunocytochemistry and Western blot after MSCs were cultured in induction medium for 7 days and 15 days.


RESULTS
MSCs differentiated into neural progenitors and expressed nestin after MSCs were incubated with medium containing IBMX for 2 d. After the medium was replaced with induction medium containing many inducing agents, MSCs differentiated into neuron-like cells and dopaminergic neuron-like cells and expressed NSE, MAP-2a, b and TH. The percentage of NSE-positive cells, MAP-2a, b-positive cells and TH-positive cells was 30.032 +/- 2.489%, 41.580 +/- 5.101% and 34.958 +/- 5.534%, respectively after MSCs were induced in medium containing GDNF, IL-1beta, mesencephalic glial-cell-conditioned medium and flash-frozen mesencephalic membrane fragments for 15 days.


CONCLUSIONS
MSCs can differentiate into dopaminergic neuron-like cells and are a new cell source for the treatment of neurodegeneration diseases and have a great potential for wide application.
Guo
Li
L
Department of Toxicology, School of Public Health, Jilin University, Changchun, Jilin 130021, China. guoli92003@yahoo.com.cn
Yin
Fei
F
Meng
Hong-Qi
HQ
Ling
Ling
L
Hu-He
Ta-Na
TN
Li
Peng
P
Zhang
Chun-Xia
CX
Yu
Shun
S
Duan
De-Sheng
DS
Fan
Hong-Xue
HX
D017667:Adipocytes / Q000166:cytology; D000818:Animals; D015153:Blotting, Western; D001854:Bone Marrow Cells; D043182:Carboxylesterase / Q000032:analysis; D002454:Cell Differentiation; D002478:Cells, Cultured; D019902:Chondrocytes / Q000166:cytology; D017077:Culture Media, Conditioned; D004298:Dopamine / Q000032:analysis; D007381:Intermediate Filament Proteins / Q000032:analysis; D008636:Mesencephalon / Q000166:cytology; D059630:Mesenchymal Stem Cells / Q000166:cytology*; D009419:Nerve Tissue Proteins / Q000032:analysis; D064231:Nestin; D009474:Neurons / Q000166:cytology* / Q000378:metabolism; D010749:Phosphoprotein Phosphatases / Q000032:analysis; D051381:Rats; D017208:Rats, Wistar
D017077:Culture Media, Conditioned; D007381:Intermediate Filament Proteins; D009419:Nerve Tissue Proteins; C577486:Nes protein, rat; D064231:Nestin; D043182:Carboxylesterase; C049301:microtubule-associated protein-2 phosphatase; D010749:Phosphoprotein Phosphatases; D004298:Dopamine