Object: To investigate the therapeutic effect of microencapsulated porcine retinal pigmented epithelial cells (RPE-M) transplantation on rat model of Parkinson's disease (PD).
Methods: Primary porcine RPE cells were harvested by enzyme digestion and expanded in culture medium. Determine the levels of dopamine (DA) and homovanillic acid (HVA) by high performance liquid chromatography electrochemical (HPLC) assay, and the levels of brain-derived neurotrophic factor (BDNF) and glial-derived neurotrophic factor (GDNF) were detected by ELISA. Alginate-polylysine-alginate (APA) microencapsulated cells were produced by using a high voltage electrostatic system. PD rat model was established by unilateral injection of 6-hydroxydopamine (6-OHDA) into the medial forebrain bundle (MFB). After that, the RPE-M was transplanted into the corpus striatum of PD rat, and then the rotation test scores were recorded and biochemical changes of the corpus striatum were tested.
Results: The levels of DA, HVA, BDNF and GDNF secreted by RPE were stable in the RPE culture supernatant and were not changed by the microencapsulation. Eighty-three percent rats developed PD by unilateral lesion of 6-OHDA in the MFB. The RPE-M transplantation had therapeutic effect on 33% PD rats.
Conclusion: Porcine RPE cells grow actively in vitro and could secrete DA, HVA, BDNF, and GDNF constantly, which does not be affected by the passage culture and the APA miroencapsulation. RPE-M transplantation of may be a curative therapy for PD.
目的: 研究微囊化后的猪视网膜色素上皮细胞(retinal pigment epithelial, RPE)对帕金森病大鼠模型的移植疗效。
方法: 原代培养 RPE 并传代, 高效液相色谱法测定培养液上清中多巴胺(dopamine, DA)和高香草酸 (homovanillic acid, HVA)的含量, ELISA 法检测脑源性神经营养因子(brain-derived neurotrophic factor, BDNF)和胶质细胞源性神经营养因子(glial-derived neurotrophic factor, GDNF)的含量。 用高压静电成囊装置制备海藻酸钠-多聚赖氨酸-海藻酸钠微囊化细胞。 6-羟基多巴胺(6-hydroxydopamine, 6-OHDA)毁损内侧前脑束 (medial fore-brain bundle, MFB)建立 SD 大鼠帕金森病模型。 立体定向移植 RPE+ 微囊, 检验旋转实验、 免疫组化和脑内生化的变化。
结果: RPE 培养上清液中DA、 HVA、 BDNF、 GDNF 的含量稳定, 微囊化后细胞长期存活, 活性没有明显变化。 6-OHDA 毁损MFB建立大鼠帕金森病模型的成模率为 83%。 移植微囊化的RPE后有效率为 33%。
结论: 猪 RPE 体外培养生长旺盛, 持续分泌 DA、 BDNF 和 GNDF, 微囊化不影响其分泌功能。 RPE 移植对帕金森病大鼠有一定的治疗作用。