全反式维甲酸促人脐血多能干细胞向多巴胺能神经元分化

山东大学学报(医学版) ›› 2012, Vol. 50 ›› Issue (6): 92-96.

全反式维甲酸促人脐血多能干细胞向多巴胺能神经元分化

李恒1，李晓红2，毕建芬3，陈亚娜4

山东大学 1.医学院，济南 250012； 2.附属济南市中心医院神经内科，济南 250013；
3.附属济南市中心医院保健五科，济南 250013； 4.附属济南市中心医院产科，济南 250013

收稿日期:2012-01-12 出版日期:2012-06-10 发布日期:2012-06-10
通讯作者: 李晓红(1960- )，女，主任医师，教授，主要从事痴呆与脑血管病的研究。 E-mail: Xiaohong_Li@sdu.edu.cn
作者简介:李恒(1983- )，男，硕士研究生，住院医师，主要从事成体干细胞的神经诱导分化的研究
基金资助:
济南市科技计划(201101104)

All-trans retinoic acid promoted human umbilical cord blood derived
multipotent stem cells to differentiate into dopaminergic neurons

LI Heng1, LI Xiao-hong2, BI Jian-fen3, CHEN Ya-na4

1. Medical School, Shandong University, Jinan 250012, China;
2. Department of Neurology, Jinan Central Hospital Affiliated to Shandong University, Jinan 250013, China;
3. Section Five, Department of Preventive Medicine, Jinan Central Hospital Affiliated to
Shandong University, Jinan 250013, China;
4. Department of Obstetrics, Jinan Central Hospital Affiliated to Shandong University, Jinan 250013, China

Received:2012-01-12 Online:2012-06-10 Published:2012-06-10

摘要/Abstract

摘要：

目的探讨在全反式维甲酸(alltrans retinoic acid, ATRA)作用下，人脐血多能干细胞(cord blood-stem cells，CB-SCs)向多巴胺能神经元的分化潜能。方法分离、培养CB-SCs后，分为诱导组、神经培养基对照组和无血清培养基对照组。诱导培养12d后，观察干细胞的形态学变化，免疫化学荧光法检测多巴胺神经元标志物的表达。ELISA法检测上清液中多巴胺递质的含量。结果 ATRA处理后，大部分CBSCs出现神经样分化，神经培养基对照组仅少量CB-SCs出现神经样分化，无血清培养基对照组CB-SCs继续扩增而无明显形态学变化；诱导组(48±11)%的细胞酪氨酸羟化酶(TH)表达阳性；神经培养基对照组仅少量TH阳性细胞；ELISA检测结果表明，诱导组多巴胺较神经培养基对照组明显升高(P<0.001)。结论 ATRA可以较为高效地促CB-SCs分化为多巴胺能神经元。

关键词: 全反式维甲酸；干细胞；脐带血；多巴胺神经元；帕金森病

Abstract:

Objective To explore the potential of human umbilical cord blood derived multipotent stem cells CB-SCs (cord blood stem cells) to differentiate into dopaminergic neurons under the treatment of all-trans retinoic acid (ATRA). Methods After isolating and culturing CB-SCs from human umbilical cord blood, stem cells were divided into three groups: the inductive group, the neuro-medium control group and the serum-free control group. After 12 days in culture, the morphology of cells was observed and dopamine neuron-associated makers were studied with immunofluorescence tests. Dopamine levels were analyzed by ELISA. Results With the treatment of ATRA, most of CB-SCs were induced into cells of typical neuronal morphology, with little change in the neuro-medium control group. Cells in the serum-free control group kept proliferating and retained morphological features of CB-SCs. As shown by immunofluorescence tests, nearly half of the cell population in the inductive group exhibited positive expression of tyrosine hydroxylase (TH). By contrast, TH+ cells in the neuromedium control group were only in small numbers. Furthermore, ELISA data established that the dopamine level was significantly higher in the inductive group relative to the neuromedium control group (P<0.001). Conclusion ATRA can efficiently induce CB-SCs to differentiate into dopamine neurons.

Key words: All-trans retinoic acid; Stem cells; Umbilical cord blood; Dopamine neuron; Parkinson disease

中图分类号:

R742.5

李恒1，李晓红2，毕建芬3，陈亚娜4. 全反式维甲酸促人脐血多能干细胞向多巴胺能神经元分化[J]. 山东大学学报(医学版), 2012, 50(6): 92-96.

LI Heng1, LI Xiao-hong2, BI Jian-fen3, CHEN Ya-na4. All-trans retinoic acid promoted human umbilical cord blood derived
multipotent stem cells to differentiate into dopaminergic neurons[J]. JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES), 2012, 50(6): 92-96.

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