CAJ | 학술논문

背景:体外培养条件下脐血间充质干细胞可向神经样细胞诱导分化.一定浓度范围的脑源性神经营养因子和睫状神经营养因子联合体外诱导可获得较高的神经元分化比例.目的:观察脑源性神经营养因子和睫状神经营养因子单独或联合体外诱导人脐带血来源的间充质干细胞分化成神经样细胞的可行性.方法:取第5代的脐血间充质干细胞,分别用5,10,20 μJ/L的脑源性神经营养因子和5,10,20 μg/L.睫状神经营养因子单独或联合诱导脐血源间充质干细胞向神经样细胞分化,另设空白对照组无任何干预措施.倒置相差显微镜观察细胞形态变化,于实验第1,3,6天分别进行神经元特异性烯醇化酶和胶质纤维酸性蛋白免疫细胞化学染色,并计数分化为神经元样细胞及神经胶质细胞的比例.结果与结论:①向神经细胞诱导后,人脐血源间充质干细胞形态明显改变,胞体收缩,胞核部分折光性增强,出现类似于树突及轴突样结构.②与空白对照组相比,脑源性神经营养因子和睫状神经营养因子能显著提高人脐血源间充质干细胞分化为神经元的比例.其中20 μg/L.的脑源性神经营养因子联合20 μg/t.睫状神经营养因子诱导人脐血源间充质干细胞分化为神经样细胞的比例最高.提示人脐血间充质干细胞经脑源性神经营养因子和睫状神经营养因子体外诱导,均能够分化为神经样细胞.
배경:체외배양조건하제혈간충질간세포가향신경양세포유도분화.일정농도범위적뇌원성신경영양인자화첩상신경영양인자연합체외유도가획득교고적신경원분화비례.목적:관찰뇌원성신경영양인자화첩상신경영양인자단독혹연합체외유도인제대혈래원적간충질간세포분화성신경양세포적가행성.방법:취제5대적제혈간충질간세포,분별용5,10,20 μJ/L적뇌원성신경영양인자화5,10,20 μg/L.첩상신경영양인자단독혹연합유도제혈원간충질간세포향신경양세포분화,령설공백대조조무임하간예조시.도치상차현미경관찰세포형태변화,우실험제1,3,6천분별진행신경원특이성희순화매화효질섬유산성단백면역세포화학염색,병계수분화위신경원양세포급신경효질세포적비례.결과여결론:①향신경세포유도후,인제혈원간충질간세포형태명현개변,포체수축,포핵부분절광성증강,출현유사우수돌급축돌양결구.②여공백대조조상비,뇌원성신경영양인자화첩상신경영양인자능현저제고인제혈원간충질간세포분화위신경원적비례.기중20 μg/L.적뇌원성신경영양인자연합20 μg/t.첩상신경영양인자유도인제혈원간충질간세포분화위신경양세포적비례최고.제시인제혈간충질간세포경뇌원성신경영양인자화첩상신경영양인자체외유도,균능구분화위신경양세포.
BACKGROUND: Umbilical cord blood mesenchymal stem cells (UCBMSCs) have been shown to differentiate into nerve-like cells under the condition of in vitro culture. Brain-derived neurotrophic factors (BDNF) combined with ciliary neurotrophic factors (CNTF) at certain concentration can in vitro induce the differentiation of UCBMSCs into high proportion of neuronal-like cells. OBJECTIVE: To investigate the feasibility of BDNF, CNTF and their combination for in vitrodifferentiation of UCBMSCs into nerve-like cells.METHODS: UCBMSCs of passage 5 were induced to differentiate into nerve-like cells using 5,10, 20 ug/L BDNF, 5,10, 20 ug/L CNTF or their combination. A blank control group was set, with no any interventions. Cell morphology was observed under inverted phase contrast microscope. At 1, 3, and 6 days of experiment, immunocytochemical staining for neuron specific enolase and glial fibrillary acidic protein was performed. The proportion of differentiated neuronal-like cells and glial cell-like cells was calculated.RESULTS ANDCONCLUSION:After induced differentiation into nerve-like cells, human UCBMSCs exhibited changeable morphology with contracted cell body, enhanced refraction of nucleus, and dendrite- and axon-like structure. Compared with blank control group, BDNF and CNTF couldsignificantly enhance the differentiation proportion of UCBMSCs into nerve-like cells. 20 ug/LBDNF combined with 20 ug/L CNTF yielded highest differentiation proportion of human UCBMSCs into nerve-like cells. These findings indicate that human UCBMSCs can differentiate into nerve-like cells after in vitro induction of BDNF, CNTF or their combination.