Department of Pediatrics, Neurobiology Program, Children's Memorial Research Center, Feinberg School of Medicine, Northwestern University, Chicago, IL 60611, USA.
Cell Transplant. 2009;18(7):801-14. doi: 10.3727/096368909X470801. Epub 2009 Sep 28.
Parkinson's disease (PD) is a common neurodegenerative disease characterized by the selective loss of dopaminergic (DA) neurons in the midbrain. Various types of stem cells that have potential to differentiate into DA neurons are being investigated as cellular therapies for PD. Stem cells also secrete growth factors and therefore also may have therapeutic effects in promoting the health of diseased DA neurons in the PD brain. To address this possibility in an experimental model of PD, bone marrow-derived neuroprogenitor-like cells were generated from bone marrow procured from healthy human adult volunteers and their potential to elicit recovery of damaged DA axons was studied in a partial lesion rat model of PD. Following collection of bone marrow, mesenchymal stem cells (MSC) were isolated and then genetically modified to create SB623 cells by transient transfection with the intracellular domain of the Notch1 gene (NICD), a modification that upregulates expression of certain neuroprogenitor markers. Ten deposits of 0.5 microl of SB623 cell suspension adjusted from 6,000 to 21,000 cells/microl in PBS or PBS alone were stereotaxically placed in the striatum 1 week after the nigrostriatal projection had been partially lesioned in adult F344 rats by injection of 6-hydroxydopamine (6-OHDA) into the striatum. At 3 weeks, a small number of grafted SB623 cells survived in the lesioned striatum as visualized by expression of the human specific nuclear matrix protein (hNuMA). In rats that received SB623 cells, but not in control rats, dense tyrosine hydroxylase immunoreactive (TH-ir) fibers were observed around the grafts. These fibers appeared to be rejuvenated host DA axons because no TH-ir in soma of surviving SB623 cells or coexpression of TH and hNuMA-ir were observed. In addition, dense serotonin immunoreactive (5-HT-ir) fibers were observed around grafted SB623 cells and these fibers also appeared to be of the host origin. Also, in some SB623 grafted rats that were sacrificed within 2 h of dl-amphetamine injection, hot spots of c-Fos-positive nuclei that coincided with rejuvenated dense TH fibers around the grafted SB623 cells were observed, suggesting increased availability of DA in these locations. Our observations suggest that NICD-transfected MSC hold potential as a readily available autologous or allogenic cellular therapy for ameliorating the degeneration of DA and 5-HT neurons in PD patients.
帕金森病（PD）是一种常见的神经退行性疾病，其特征是中脑中多巴胺能（DA）神经元的选择性丧失。各种类型的具有分化为 DA 神经元潜力的干细胞被用作 PD 的细胞治疗。干细胞还分泌生长因子，因此也可能具有促进 PD 大脑中患病 DA 神经元健康的治疗作用。为了在 PD 的实验模型中解决这种可能性，从健康的成年人类志愿者中采集骨髓，生成骨髓源性神经祖细胞样细胞，并在 PD 的部分损伤大鼠模型中研究其诱导受损 DA 轴突恢复的潜力。收集骨髓后，分离间充质干细胞（MSC），然后通过瞬时转染 Notch1 基因（NICD）的细胞内域对其进行基因修饰，从而创建 SB623 细胞，该修饰上调某些神经祖细胞标志物的表达。将浓度为 6000 至 21000 个细胞/微升的 0.5 微升 SB623 细胞悬液的 10 个沉积点以立体定向方式置于纹状体中，1 周后，通过将 6-羟多巴胺（6-OHDA）注入纹状体对成年 F344 大鼠的黑质纹状体投射进行部分损伤。在 3 周时，通过表达人类特异性核基质蛋白（hNuMA），可以在受损的纹状体中观察到少量存活的 SB623 细胞。在接受 SB623 细胞的大鼠中，但在对照大鼠中，观察到围绕移植物的密集酪氨酸羟化酶免疫反应性（TH-ir）纤维。这些纤维似乎是再生的宿主 DA 轴突，因为在存活的 SB623 细胞的体中未观察到 TH-ir 或 TH 和 hNuMA-ir 的共表达。此外，在移植的 SB623 细胞周围观察到密集的血清素免疫反应性（5-HT-ir）纤维，这些纤维似乎也来自宿主。此外，在一些在 dl-安非他命注射后 2 小时内被处死的 SB623 移植大鼠中，观察到与移植的 SB623 细胞周围再生的密集 TH 纤维重合的 c-Fos 阳性核热点，表明这些部位 DA 的可用性增加。我们的观察结果表明，NICD 转染的 MSC 具有作为一种现成的自体或同种异体细胞疗法的潜力，可改善 PD 患者的 DA 和 5-HT 神经元变性。
