Understanding microchannel culture: parameters involved in soluble factor signaling
Understanding microchannel culture: parameters involved in soluble factor signaling
复制标题
DOI：
10.1039/b618793e
复制
发表时间：
2007-06-01
期刊：
影响因子：
6.1
通讯作者：
Beebe, David J.
中科院分区：
文献类型：
--
作者：
Yu, Hongmei;Alexander, Caroline M.;Beebe, David J.
虽然体内自分泌-旁分泌信号传导的重要性是清楚的，但研究体外分泌的内源性因子的作用的能力受到典型培养平台的阻碍。在多孔板中，大的空气-液体界面引起对流流动，该对流流动持续地混合流体，破坏局部基于扩散的积聚。简单的微通道提供了一个更可控的微环境，可用于研究分泌因子的影响。在这里，我们利用微通道培养检查基本的培养参数和它们之间的相互作用，使用正常的乳腺上皮细胞（NMuMG）。研究了以下参数：（1）细胞密度（80 vs. 240个细胞mm（-2）），（2）外源生长因子（表皮生长因子[EGF] vs.胎牛血清），（3）培养基更换频率（1 It，4 It，12 h），和（4）培养平台（微通道vs. 96孔板）。与96孔板相比，细胞在微通道中表现出增加的生长速率。随着培养基更换频率的降低，细胞增殖增加。对于所使用的微通道几何形状，在几个小时内达到重要的阈值浓度。总之，结果表明，四个因素的功能和它们之间的相互作用对NMuMG生长的空间和时间相关的分子扩散在受控的微通道空间。无对流的微通道环境可能被证明是有用的研究可溶性因子信号在体外，并测试模型和预测的自分泌-旁分泌信号。
While the importance of autocrine-paracrine signaling in vivo is clear, the ability to study the effects of secreted endogenous factors in vitro is hampered by canonical culture platforms. In multi-well plates, the large air-liquid interface gives rise to convective flows that continually mix the fluid disrupting the local diffusion-based accumulation. Simple microchannels provide a more controlled microenvironment that can be used to study secreted factor effects. Here, we utilize microchannel culture to examine basic culture parameters and their interactions using normal mammary gland epithelial cells (NMuMG). The following parameters were studied: (1) cell density (80 vs. 240 cells mm(-2)), (2) exogenous growth factors (epidermal growth factor [EGF] vs. fetal bovine serum), (3) medium change frequency (I It, 4 It, 12 h), and (4) culture platform (microchannels vs. 96-well plates). The cells exhibited increased growth rates in microchannels as compared to 96-well plates. Cell proliferation increased as the frequency of media change decreased. For the microchannel geometries used, important threshold concentrations were reached in a few hours. In aggregate, the results indicate that the function of the four factors and their interactions on NMuMG growth are spatially and temporally related by molecular diffusion in the controlled microchannel space. The convective-free microchannel environment may prove useful for studying soluble factor signaling in vitro, and to test models and predictions of autocrine-paracrine signaling.