Department of Systems Neuroscience, Tokyo Medical and Dental University Graduate School of Medical and Dental Sciences, Yushima, Bunkyo-ku, Japan.
Biochem J. 2010 Mar 29;427(2):297-304. doi: 10.1042/BJ20100071.
Attachment to the substrate is essential for both survival and differentiation of various kinds of cells, such as neurons and epithelial cells. We recently found a small synthetic molecule, adhesamine, which boosts adhesion and growth of mammalian cells. In the present study, we applied adhesamine to primary cultured hippocampal neuronal cells and compared its effects with those of PLL (poly-L-lysine), which is widely used as a substrate for cell cultures. Neurons grown on adhesamine-coated coverslips survived for up to 1 month without a feeder layer of glial cells, and had greater viability than cells grown on PLL-coated coverslips. Morphological analysis revealed that neurons cultured with adhesamine exhibited earlier differentiation, i.e. earlier axonal outgrowth and dendritic maturation with enhanced neurite branching, than neurons cultured with PLL. Synaptic formation and postsynaptic responses were evident as early as 4 days in cells cultured with adhesamine. Acceleration of differentiation is mediated by earlier activation of the signalling pathways from heparan sulfate in the extracellular matrix to both FAK (focal adhesion kinase) and MAPK (mitogen-activated protein kinase). Improved survival rates and accelerated maturation of neurons exposed to adhesamine suggest that this completely synthetic molecule may be a useful reagent for culturing neuronal cells.
细胞黏附对于各种细胞（如神经元和上皮细胞）的存活和分化是必不可少的。我们最近发现了一种小分子合成物，黏附素，它能促进哺乳动物细胞的黏附和生长。在本研究中，我们将黏附素应用于原代培养的海马神经元细胞，并将其与广泛用作细胞培养底物的多聚赖氨酸（PLL）进行比较。在黏附素涂层盖玻片上生长的神经元在没有胶质细胞饲养层的情况下可存活长达 1 个月，且比在 PLL 涂层盖玻片上生长的细胞具有更高的存活率。形态分析表明，用黏附素培养的神经元表现出更早的分化，即更早的轴突生长和树突成熟，具有增强的神经突分支，而用 PLL 培养的神经元则没有。在用黏附素培养的细胞中，早在第 4 天就可以观察到突触形成和突触后反应。分化的加速是通过细胞外基质中硫酸乙酰肝素与粘着斑激酶（FAK）和丝裂原活化蛋白激酶（MAPK）之间信号通路的早期激活来介导的。黏附素处理的神经元的存活率提高和成熟加速表明，这种完全合成的分子可能是培养神经元的有用试剂。
