骨髓间充质干细胞移植入帕金森大鼠模型的纹状体后分化为多巴胺分泌细胞

目的探讨大鼠骨髓间充质干细胞(rMSCs)向多巴胺能神经元分化条件及其体内修复大鼠帕金森模型的潜能。方法为了能够更好修复大鼠单侧下丘脑-黑质部位注射6-OHDA所造成的功能毁损,我们从SD大鼠中提取了rMSCs。首先利用中药麝香多肽-1诱导rMSCs在体外无血清培养基中扩增并分化为神经元样细胞,然后移植到单侧纹状体区DA耗竭的大鼠模型中。结果加入麝香多肽-1后2 h,细胞开始分化成神经元样细胞,细胞团呈NSE及NF-H阳性。移植后动物模型较对照组有明显的功能改善(P<0．001),表现为25只实验鼠阿普吗啡诱导的旋转减轻,平均持续56 d。免疫组织化学分析证实大量来自rMSCs的细胞在移植带存活并可迁移到距损伤处5 mm的部位,通过检测多巴胺能神经元特异性标志TH发现大约50％～55%细胞继续向多巴胺能神经元方向分化。结论以上发现说明rMSCs来源的细胞移植到大鼠纹状体区后能够存活、迁移、并自发向多巴胺分泌细胞分化,并能修复源自于中脑多巴胺能神经元的功能。

Functional differentiation of MSCs into dopamine-producing cells after engrafting into the striatum of Parkinson''''s rat

Objective To investigate the factors of rMSCs differentiation into dopamine-producing cells and its recovery potential of rat Parkinson model in vivo. Methods To better repair the 6-hydroxydopamine (6-OHDA) unilateral lesioned dysfunction in the substantia nigra of rats, rMSCs was prepared from adult Sprague-Dawley (SD) rat. They were firstly proliferated and differentiated in vitro into neuron-like cells in serum-free medium with a Chinese medicine, Musk's polypeptide named Musk-1, then grafted into Parkinson rats that had unilateral Dopaminergic (DA) - depleted striatum. Results 2 h after Musk-1 was added, cells started to differentiate into neuron-like cells, making those cell spheres nestin, NSE, NF-H positive. After cell implantation, animals promoted significant improvement in function (apomorphine-induced rotation ameliorated in 25 grafted animals and persisted for 56 days on average) compared to the lesion-control group (P<.001). Histological analysis confirmed a large number of rMSCs-derived cells survived well in the transplantation zoon and migrated around the host lesion as far as 5 millimeters, followed by developing into DA neuron phenotype on approximately 50%~55% of overall cells tested by the DA neuron marker tyrosine hydroxylase (TH). Conclusion These findings demonstrated that grafted rMSCs-derived cells can survive, migrate, and spontaneously develop into Dopamine-producing cells in striatum and restore behavioral properties of neurons expected from the midbrain.