Kim Jin Hee, Sun Woong, Han Dong Wook, Lim Dong-Jun, Lee Jangbo
Department of Neurosurgery, College of Medicine, Korea University, Seoul, Korea.
Neurol Sci. 2015 Apr;36(4):527-34. doi: 10.1007/s10072-014-2012-1. Epub 2014 Nov 20.
Reprogramming of fibroblasts into induced neural stem cells (NSCs) is a potentially unlimited source of neurons. In this study, we cocultured cortical neuronal cells with iNSCs in a transwell system. We then investigated the effects of coculture on apoptosis and the secretion of cytokines and growth factors. Compared with the cultured cortical neuronal culture alone, cortical neuronal cells cocultured with iNSCs exhibited increased proliferation. TUNEL assay was used to assess the rate of apoptosis at selected time intervals (24, 48 and 72 h). Cells cocultured with iNSCs had fewer apoptotic cells than those cultured without iNSCs. When TUNEL assay was performed in parallel with staining for the neuronal marker Tuj1, the number of neuronal apoptotic cells was found to be lower in cells cocultured with iNSCs than in those cultured without iNSCs for 72 h. Secretion of cytokines and growth factors by iNSCs was evaluated by ELISA. Compared to cells cultured without iNSCs, coculture decreased levels of the inflammatory cytokines and increased levels of HGF and VEGF. These findings indicated that iNSCs could be used as a new treatment strategy for neurodegenerative conditions by promoting proliferation and decreasing apoptosis of cortical neuronal cells.
将成纤维细胞重编程为诱导神经干细胞（NSCs）是一种潜在的无限神经元来源。在本研究中，我们在Transwell系统中将皮质神经元细胞与诱导神经干细胞共培养。然后，我们研究了共培养对细胞凋亡以及细胞因子和生长因子分泌的影响。与单独培养的皮质神经元相比，与诱导神经干细胞共培养的皮质神经元细胞增殖增加。使用TUNEL法在选定的时间间隔（24、48和72小时）评估细胞凋亡率。与未与诱导神经干细胞共培养的细胞相比，与诱导神经干细胞共培养的细胞凋亡细胞更少。当TUNEL法与神经元标志物Tuj1染色同时进行时，发现与诱导神经干细胞共培养72小时的细胞中神经元凋亡细胞的数量低于未与诱导神经干细胞共培养的细胞。通过ELISA评估诱导神经干细胞分泌细胞因子和生长因子的情况。与未与诱导神经干细胞共培养的细胞相比，共培养降低了炎性细胞因子的水平，并增加了HGF和VEGF的水平。这些发现表明，诱导神经干细胞可通过促进皮质神经元细胞的增殖和减少其凋亡，用作神经退行性疾病的一种新的治疗策略。
