Transform human umbilical cord blood cell into dopaminergic neurons

• Main Authors: SHIH HAO MING, 石濠銘
• Other Authors: 馬國興
• Format: Others
• Language: zh-TW
• Published: 2006
• Online Access: http://ndltd.ncl.edu.tw/handle/43935582774272147052

碩士 === 國防醫學院 === 生物及解剖學研究所 === 94 === A cluster of cells that can secret dopamine as neurotransmitter refer to dopaminergic neurons. The severe loss of the dopaminergic neurons gives rise to the dopamine deficiency in the nigrostriatal system causing progressive degenerative disorder called Parkinson’s disease. In this experiment we expect human umbilical cord blood (HUCB) may differentiate into neuroepithelial cells by manipulation, and we expect that these cells might transform into specific neuron type such as dopaminergic neurons. In line with this hypothesis, we cultured mononucleocyte layer cells of HUCB separated by Ficoll-Paque Plus in 10ng/ml EGF and 10ng/ml bFGF contented MEM with 20% FBS for three days and then subcultured with 0.25μM retinoic acid, 1μM 2-mercaptoethanol, and 10ng/ml brain-derived growth factor to induce for six days. After the treatment, we found nestin positive cells (neuroepithelial progenitor cell) counting up to 4.87-17.73% of total cells in adhere layer, and there were cells β-tubulin III, microtubule associated protein 2, and tyrosine hydroxylase positive cells in the cultures. The western blot study also confirm TH expression in the cultures. These finding encourage us to transplant these induced cells into the striatum of hemiparkisonian rats. In the amphetamine-inducing rotation behavior test, there were no changes for rotational recovery. The immunohistochemistry study demonstrate that there were human cells and TH positive cells existed in the grafted area. The data of this experiment may provide another therapeutic possibility for Parkinson’s disease.