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Abstract

BACKGROUND:

Parkinson's disease treated by cell replacement therapy has become a hot research at present, and how to obtain adequate and effective dopaminergic neurons in vitro is a strategy in the treatment of the disease.

OBJECTIVE:

To investigate the feasibility of human umbilical cord blood-derived mesenchymal stem cells differentiating into dopaminergic neurons in vitro.

METHODS:

The fifth passage human umbilical cord blood-derived mesenchymal stem cells were inoculated with 5×103/mL. The cells were pre-inducted with 20 ng/mL epidermal growth factor+20 ng/mL basic fibroblast growth factor for 24 hours. Then the cells were divided into four groups: the control group was not given induction medium but only cultured in DMEM/F12 culture medium containing 10% fetal bovine serum; the other three induced groups were induced with 100 μmol/L ascorbic acid, 1 μmol/L all-trans retinoic acid 50 μg/mL glial cell line-derived neurotrophic factor alone or in combination respectively.

RESULTS AND CONCLUSION:

The expression of tyrosine hydroxylase, dopamine transporter and dopamine receptor D2 mRNA could be seen in the induced groups. After induction and differentiation, the cells could express the neurons and glial cell-specific antigens. Compared with the control group, the positive rates of nestin, neuron-specific enolase, glial fibrillary acidic protein, tyrosine hydroxylase, dopamine transporter and dopamine receptor D2 in the induced groups were significantly increased (P < 0.05); the positive rates in the all-trans retinoic acid+glial cell line-derived neurotrophic factor group and the combination induced group were significantly higher than those in the ascorbic acid group (P < 0.05), but the increased degree of the combination induced group was significantly higher than that of the all-trans retinoic acid+glial cell line-derived neurotrophic factor group (P < 0.05). Ascorbic acid, all-trans retinoic acid+glial cell line-derived neurotrophic factor, all-trans retinoic acid+glial cell line-derived neurotrophic factor+ascorbic acid can promote the human umbilical cord blood-derived mesenchymal stem cells to differentiate into dopaminergic neurons, and the combined induction can get the best effect.

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