Effects of substrate stiffness and cell density on primary hippocampal cultures
Effects of substrate stiffness and cell density on primary hippocampal cultures
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DOI：
10.1016/j.jbiosc.2010.04.004
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发表时间：
2010-10-01
影响因子：
2.8
通讯作者：
Firestein, Bonnie L.
中科院分区：
文献类型：
--
作者：
Previtera, Michelle L.;Langhammer, Christopher G.;Firestein, Bonnie L.
以前的研究表明，树突的影响，基板刚度时，神经元接种在纯或混合培养。然而，由于基板刚度也可以影响其他方面的文化发展的影响树突分支，如总细胞数，目前还不清楚是否基板刚度施加直接或间接影响树突形态。在这项研究中，我们确定基板刚度是否起着至关重要的作用，在调节独立的细胞数量的树突分支。我们将原代混合海马培养物分别接种在软凝胶和硬凝胶上，杨氏模量分别为1 kPa和7 kPa。我们发现，在相同的细胞数量下，相对于生长在较软基质上的神经元，接种在较硬基质上的神经元表现出增加的分支。在硬凝胶上，我们还观察到细胞数量依赖性效应，其中增加初始电镀密度降低树突分支。这种变化与细胞外谷氨酸的增加相关。我们的结论是，细胞数量和基板刚度发挥作用，在确定枝晶分支，这两个效果是相互独立的。(C)2010年，生物技术学会，日本。All rights reserved.
Previous studies have shown that dendrites are influenced by substrate stiffness when neurons are plated in either pure or mixed cultures. However, because substrate rigidity can also affect other aspects of culture development known to impact dendrite branching, such as overall cell number, it is unclear whether substrate stiffness exerts a direct or indirect effect on dendrite morphology. In this study, we determine whether substrate stiffness plays a critical role in regulating dendrite branching independent of cell number. We plated primary mixed hippocampal cultures on soft and stiff gels, with Young's moduli of 1 kPa and 7 kPa, respectively. We found that neurons plated on stiffer substrates showed increased branching relative to neurons grown on softer substrates at the same cell number. On the stiff gels, we also observed a cell number-dependent effect, in which increasing initial plating density decreased dendrite branching. This change correlates with an increase in extracellular glutamate. We concluded that both cell number and substrate stiffness play roles in determining dendrite branching, and that the two effects are independent of one another. (C) 2010, The Society for Biotechnology, Japan. All rights reserved.