Control protocol for robust in vitro glial scar formation around microwires: essential roles of bFGF and serum in gliosis.
Control protocol for robust in vitro glial scar formation around microwires: essential roles of bFGF and serum in gliosis.
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DOI：
10.1016/j.jneumeth.2009.05.002
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发表时间：
2009-07-30
影响因子：
3
通讯作者：
Reichert, William M.
中科院分区：
文献类型：
--
作者：
Polikov, Vadim S.;Su, Eric C.;Ball, Matthew A.;Hong, Jau-Shyong;Reichert, William M.
以前，我们报道了一种体外细胞培养模型，该模型重现了用于记录大脑的电极周围神经胶质瘢痕的许多特征;然而，该模型缺乏建立有用的表征工具所需的重现性。该方法描述了一种方案，该方案以通常用于培养神经干细胞/前体细胞的方案为模型，产生可预测的强烈瘢痕形成反应的阳性对照。系统地改变六个独立的细胞培养变量（生长培养基、接种密度、bFGF添加日、处理培养基中的血清浓度、处理日和培养持续时间），并对所得瘢痕进行定量。发现以下条件产生最高水平的瘢痕形成：补充有B27的Neurobasal培养基，处理时10%胎牛血清，在接种和处理时添加10 ng/ml b-FGF，接种后至少6天处理，瘢痕生长至少5天。只要每孔至少接种500，000个细胞，接种密度就不会影响瘢痕形成，但适当的培养基、bFGF和血清对于显著的瘢痕形成至关重要-这些见解有助于验证基于体外的方法来理解神经胶质瘢痕形成。由于本研究中开发的对照方案在每次解剖时均产生强的、可重现的胶质瘢痕形成阳性对照，因此该培养模型适用于胶质瘢痕形成和神经电极失效机制的体外研究。
Previously, we reported an in vitro cell culture model that recreates many of the hallmarks of glial scarring around electrodes used for recording in the brain; however, the model lacked the reproducibility necessary to establish a useful characterization tool. This methods paper describes a protocol, modeled on protocols typically used to culture neural stem/precursor cells, that generates a predictable positive control of an intense scarring reaction. Six independent cell culture variables (growth media, seeding density, bFGF addition day, serum concentration in treatment media, treatment day, and duration of culture) were varied systematically and the resulting scars were quantified. The following conditions were found to give the highest level of scarring: Neurobasal medium supplemented with B27, 10% fetal bovine serum at treatment, 10ng/ml b-FGF addition at seeding and at treatment, treatment at least 6 days after seeding and scar growth of at least 5 days. Seeding density did not affect scarring as long as at least 500,000 cells were seeded per well, but appropriate media, bFGF, and serum were essential for significant scar formation - insights that help validate the in vitro-based approach to understanding glial scarring. With the control protocol developed in this study producing a strong, reproducible glial scarring positive control with every dissection, this culture model is suitable for the in vitro study of the mechanisms behind glial scarring and neuroelectrode failure.
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影响因子：
4.2
作者：
Magnus, Tim;Coksaygan, Turhan;Mattson, Mark P.
通讯作者：
Mattson, Mark P.
影响因子：
14
作者：
Polikov, Vadim S.;Block, Michelle L.;Reichert, W. Monty
通讯作者：
Reichert, W. Monty
DOI：
10.1016/0169-328x(95)00318-m
发表时间：
1996-04-01
期刊：
MOLECULAR BRAIN RESEARCH
影响因子：
--
作者：
Pennypacker, KR;Hong, JS;McMillian, MK
通讯作者：
McMillian, MK
DOI：
10.1109/86.788468
发表时间：
1999-09-01
期刊：
IEEE transactions on rehabilitation engineering : a publication of the IEEE Engineering in Medicine and Biology Society
影响因子：
--
作者：
Liu, X;McCreery, D B;Agnew, W F
通讯作者：
Agnew, W F
影响因子：
64.8
作者：
Hermanson, O;Jepsen, K;Rosenfeld, MG
通讯作者：
Rosenfeld, MG