DAS Mainak, Bhargava Neelima, Gregory Cassie, Riedel Lisa, Molnar Peter, Hickman James J
Nanoscience Technology Center, University of Central Florida, Orlando, Florida 32826, USA.
In Vitro Cell Dev Biol Anim. 2005 Nov-Dec;41(10):343-8. doi: 10.1007/s11626-005-0006-2.
In this study, we have documented by morphological analysis, immunocytochemistry, and electrophysiology, the development of a culture system that promotes the growth and long-term survival of dissociated adult rat spinal cord neurons. This system comprises a patternable, nonbiological, cell growth-promoting organosilane substrate coated on a glass surface and an empirically derived novel serum-free medium, supplemented with specific growth factors (acidic fibroblast growth factor, heparin sulfate, neurotrophin-3, brain-derived neurotrophic factor, glial-derived neurotrophic factor, cardiotrophin-1, and vitronectin). Neurons were characterized by immunoreactivity for neurofilament 150, neuron-specific enolase, Islet-1 antibodies, electrophysiology, and the cultures were maintained for 4-6 wk. This culture system could be a useful tool for the study of adult mammalian spinal neurons in a functional in vitro system.
在本研究中,我们通过形态学分析、免疫细胞化学和电生理学方法,记录了一种促进成年大鼠脊髓解离神经元生长和长期存活的培养系统的建立。该系统包括涂覆在玻璃表面的可图案化、非生物的细胞生长促进有机硅烷底物,以及经验性得出的新型无血清培养基,补充有特定生长因子(酸性成纤维细胞生长因子、硫酸肝素、神经营养因子-3、脑源性神经营养因子、胶质细胞源性神经营养因子、心肌营养素-1和玻连蛋白)。通过对神经丝150、神经元特异性烯醇化酶、胰岛-1抗体的免疫反应性以及电生理学对神经元进行表征,并且将培养物维持4-6周。这种培养系统可能是在功能性体外系统中研究成年哺乳动物脊髓神经元的有用工具。