The effect of the controlled release of nerve growth factor from collagen gel on the efficiency of neural cell culture
The effect of the controlled release of nerve growth factor from collagen gel on the efficiency of neural cell culture
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DOI：
10.1016/j.biomaterials.2008.09.021
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发表时间：
2009-01-01
期刊：
影响因子：
14
通讯作者：
Kim, Byung-Soo
中科院分区：
文献类型：
--
作者：
Bhang, Suk Ho;Lee, Tae-Jin;Kim, Byung-Soo
在这项研究中，我们测试的假设，即嗜铬细胞瘤（PC 12）细胞培养所需的神经生长因子（NGF）的量可以显着减少控制释放的神经生长因子从胶原蛋白凝胶涂层的培养表面上。将细胞在负载有不同量的NGF的胶原凝胶上培养。作为对照，在胶原凝胶上培养PC 12细胞，每天向培养基中加入不同量的NGF。在最初的12小时爆发后，NGF从凝胶中稳定释放4天。分别使用末端尿苷缺口末端标记和3-（4，5-二甲基噻唑-2-基）-2，5-二苯基溴化四唑测定来测定凋亡活性和细胞活力。免疫细胞化学和Western blot分析确定神经元分化。与每天添加100 ng NGF（3天内添加300 ng）相比，每天添加10 ng NGF显示细胞活力和神经元分化显著降低，凋亡活性增加。与此相反，胶原凝胶加载10 ng的神经生长因子产生细胞活力，凋亡活性，和神经元分化与100 ng的神经生长因子每天添加的文化相似。我们的方法将PC 12细胞培养所需的NGF量减少到每日添加量的1/3，而不影响细胞活力、凋亡或分化。这种方法可以通过减少所需的昂贵生长因子的量来节省干细胞的大规模培养。(C)2008爱思唯尔有限公司保留所有权利。
In this study, we tested the hypothesis that the amount of nerve growth factor (NGF) required for pheochromocytoma (PC12) cell culture can be dramatically reduced by controlled release of NGF from a collagen gel coating on the culture surface. Cells were cultured on collagen gels loaded with various amounts of NGF. As a control, PC12 cells were cultured on collagen gels with daily addition of various amounts of NGF to the culture medium. After an initial 12 h burst, NGF was steadily released from the gels for 4 days. Apoptotic activity and cell viability were determined using terminal uridine nick end labeling and the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay, respectively. Neuronal differentiation was determined using immunocytochemistry and Western blot analysis. Compared to 100 ng NGF daily addition (300 ng over 3 days), 10 ng NGF daily addition showed dramatically decreased cell viability and neuronal differentiation and increased apoptotic activity. In contrast, collagen gels loaded with 10 ng NGF yielded cell viability, apoptotic activity, and neuronal differentiation similar to those of culture with 100 ng NGF daily addition. Our method reduced the amount of NGF required for PC12 cell culture to 1/3th of that used in daily addition without affecting cell viability, apoptosis, or differentiation. This method could economize large-scale culture of stem cells by reducing the amount of costly growth factors needed. (C) 2008 Elsevier Ltd. All rights reserved.