School of Science for Open and Environmental Systems, Graduate School of Science and Technology, Keio University, Yokohama, Kanagawa, Japan.
Department of Mechanical Engineering, Faculty of Science and Technology, Keio University, Yokohama, Kanagawa, Japan.
PLoS One. 2020 Jul 15;15(7):e0235827. doi: 10.1371/journal.pone.0235827. eCollection 2020.
Homogenization of the initial cell distribution is essential for effective cell development. However, there are few previous reports on efficient cell seeding methods, even though the initial cell distribution has a large effect on cell proliferation. Dense cell regions have an inverse impact on cell development, known as contact inhibition. In this study, we developed a method to homogenize the cell seeding density using secondary flow, or Ekman transportation, induced by orbital movement of the culture dish. We developed an orbital shaker device that can stir the medium in a 35-mm culture dish by shaking the dish along a circular orbit with 2 mm of eccentricity. The distribution of cells in the culture dish can be controlled by the rotational speed of the orbital shaker, enabling dispersion of the initial cell distribution. The experimental results indicated that the cell density became most homogeneous at 61 rpm. We further evaluated the cell proliferation after homogenization of the initial cell density at 61 rpm. The results revealed 36% higher proliferation for the stirred samples compared with the non-stirred control samples. The present findings indicate that homogenization of the initial cell density by Ekman transportation contributes to the achievement of higher cell proliferation.
细胞初始分布的均化对于有效的细胞发育至关重要。然而，尽管初始细胞分布对细胞增殖有很大影响，但之前很少有关于高效细胞接种方法的报道。密集的细胞区域对细胞发育有相反的影响，称为接触抑制。在这项研究中，我们开发了一种使用培养皿轨道运动引起的二次流或 Ekman 输运来均匀化细胞接种密度的方法。我们开发了一种轨道摇床装置，通过沿偏心 2 毫米的圆形轨道摇动培养皿，可在 35-mm 培养皿中搅拌培养基。通过轨道摇床的转速可以控制培养皿中细胞的分布，从而实现初始细胞分布的分散。实验结果表明，在 61rpm 时细胞密度变得最均匀。我们进一步评估了在 61rpm 时初始细胞密度均化后的细胞增殖情况。结果表明，与未搅拌的对照样品相比，搅拌样品的增殖率高 36%。本研究结果表明，Ekman 输运引起的初始细胞密度均化有助于实现更高的细胞增殖。
