Naghdi Majid, Tiraihi Taki, Namin Seyed Alireza Mesbah, Arabkheradmand Jalil
Department of Anatomical Sciences, Tarbiat Modares University, Tehran, Iran.
Cytotherapy. 2009;11(2):137-52. doi: 10.1080/14653240802716582.
Cholinergic neurons are very important cells in spinal cord injuries because of the deficits in motor, autonomic and sensory neurons. In this study, bone marrow stromal cells (BMSC) were evaluated as a source of cholinergic neurons in a rat model of contusive spinal cord injury.
BMSC were isolated from adult rats and transdifferentiated into cholinergic neuronal cells. The BMSC were pre-induced with beta-mercaptoethanol (BME), while the induction was done with nerve growth factor (NGF). Neurofilament (NF)-68, -160 and -200 immunostaining was used for evaluating the transdifferentiation of BMSC into a neuronal phenotype. NeuroD expression, a marker for neuroblast differentiation, and Oct-4 expression, a marker for stemness, were evaluated by reverse transcriptase (RT)-polymerase chain reaction (PCR). Choline acetyl transferase (ChAT) immunoreactivity was used for assessing the cholinergic neuronal phenotype. Anti-microtubule-associated protein-2 (MAP-2) and anti-synapsin I antibodies were used as markers for the tendency for synptogenesis. Finally, the induced cells were transplanted into the contused spinal cord and locomotion was evaluated with the Basso-Beattie-Bresnahan (BBB) test.
At the induction stage, there was a decline in the expression of NF-68 associated with a sustained increase in the expression of NF-200, NF-160, ChAT and synapsin I, whereas MAP-2 expression was variable. Transplanted cells were detected 6 weeks after their injection intraspinally and were associated with functional recovery.
The transdifferentiation of BMSC into a cholinergic phenotype is feasible for replacement therapy in spinal cord injury.
由于运动、自主神经和感觉神经元的功能缺陷，胆碱能神经元在脊髓损伤中是非常重要的细胞。在本研究中，在大鼠挫伤性脊髓损伤模型中评估了骨髓基质细胞（BMSC）作为胆碱能神经元来源的情况。
从成年大鼠中分离出BMSC并将其转分化为胆碱能神经元细胞。BMSC先用β-巯基乙醇（BME）进行预诱导，而诱导则用神经生长因子（NGF）进行。使用神经丝（NF）-68、-160和-200免疫染色来评估BMSC向神经元表型的转分化。通过逆转录（RT）-聚合酶链反应（PCR）评估神经母细胞分化标志物NeuroD的表达和干性标志物Oct-4的表达。使用胆碱乙酰转移酶（ChAT）免疫反应性来评估胆碱能神经元表型。使用抗微管相关蛋白-2（MAP-2）和抗突触素I抗体作为突触形成倾向的标志物。最后，将诱导的细胞移植到挫伤的脊髓中，并用Basso-Beattie-Bresnahan（BBB）试验评估运动能力。
在诱导阶段，NF-68的表达下降，同时NF-200、NF-160、ChAT和突触素I的表达持续增加，而MAP-2的表达则有所不同。脊髓内注射后6周检测到移植细胞，且与功能恢复相关。
BMSC向胆碱能表型的转分化对于脊髓损伤的替代治疗是可行的。
