Growth of primary embryo cells in a microculture system.
Growth of primary embryo cells in a microculture system.
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原代胚胎细胞在微培养系统中的生长。
DOI：
10.1007/s10544-009-9380-y
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发表时间：
2010
影响因子：
2.8
通讯作者：
Fan,Tai-Hsi
中科院分区：
文献类型：
--
作者：
Villa,Max;Pope,Sara;Conover,Joanne;Fan,Tai-Hsi
我们提出了最佳的灌注条件下生长的原代小鼠胚胎成纤维细胞（mEFs）和小鼠胚胎干细胞（mESCs）使用微流控灌注培养系统。为了平衡营养更新，同时确保细胞分泌因子的存在，我们发现在我们的实验设置中培养原代胚胎成纤维细胞（mEFs）的最佳灌注速率为10 nL/min，微通道中的平均流速为0.55 μm/s。原代mEFs可能有更大的依赖于细胞分泌的因子相比，他们的永生对应3 T3成纤维细胞在类似的条件下培养。接种密度和灌注速率对原代细胞的增殖都是至关重要的。使用微培养系统培养mEF和mESC一周，表现出与培养皿中生长的那些相似的形态和活力。使用荧光免疫测定法分析mEFs和mESCs，以确定其增殖状态和蛋白表达。我们的研究结果表明，基于灌注的微培养环境能够支持多能胚胎干细胞的高度增殖状态。
We present optimal perfusion conditions for the growth of primary mouse embryonic fibroblasts (mEFs) and mouse embryonic stem cells (mESCs) using a microfluidic perfusion culture system. In an effort to balance nutrient renewal while ensuring the presence of cell secreted factors, we found that the optimal perfusion rate for culturing primary embryonic fibroblasts (mEFs) in our experimental setting is 10 nL/min with an average flow velocity 0.55 μm/s in the microchannel. Primary mEFs may have a greater dependence on cell secreted factors when compared to their immortalized counterpart 3T3 fibroblasts cultured under similar conditions. Both the seeding density and the perfusion rate are critical for the proliferation of primary cells. A week long cultivation of mEFs and mESCs using the microculture system exhibited similar morphology and viability to those grown in a petri dish. Both mEFs and mESCs were analyzed using fluorescence immunoassays to determine their proliferative status and protein expression. Our results demonstrate that a perfusion-based microculture environment is capable of supporting the highly proliferative status of pluripotent embryonic stem cells.