Kagawa Yuki, Kino-Oka Masahiro
Department of Biotechnology, Graduate School of Engineering , Osaka University , 2-1 Yamada-oka, Suita, Osaka 565-0871 , Japan.
R Soc Open Sci. 2016 Oct 26;3(10):160500. doi: 10.1098/rsos.160500. eCollection 2016 Oct.
Regenerative therapy using autologous skeletal myoblasts requires a large number of cells to be prepared for high-level secretion of cytokines and chemokines to induce good regeneration of damaged regions. However, myoblast expansion culture is hindered by a reduction in growth rate owing to cellular quiescence and differentiation, therefore optimization is required. We have developed a kinetic computational model describing skeletal myoblast proliferation and differentiation, which can be used as a prediction tool for the expansion process. In the model, myoblasts migrate, divide, quiesce and differentiate as observed during culture. We assumed cell differentiation initiates following cell-cell attachment for a defined time period. The model parameter values were estimated by fitting to several predetermined experimental datasets. Using an additional experimental dataset, we confirmed validity of the developed model. We then executed simulations using the developed model under several culture conditions and quantitatively predicted that non-uniform cell seeding had adverse effects on the expansion culture, mainly by reducing the existing ratio of proliferative cells. The proposed model is expected to be useful for predicting myoblast behaviours and in designing efficient expansion culture conditions for these cells.
使用自体骨骼肌成肌细胞的再生疗法需要准备大量细胞,以便高水平分泌细胞因子和趋化因子,从而诱导受损区域良好再生。然而,由于细胞静止和分化导致生长速率降低,成肌细胞的扩增培养受到阻碍,因此需要进行优化。我们开发了一个动力学计算模型,用于描述骨骼肌成肌细胞的增殖和分化,该模型可作为扩增过程的预测工具。在该模型中,成肌细胞在培养过程中会发生迁移、分裂、静止和分化。我们假设细胞分化在细胞间附着一段特定时间后开始。通过拟合几个预先确定的实验数据集来估计模型参数值。使用一个额外的实验数据集,我们证实了所开发模型的有效性。然后,我们在几种培养条件下使用所开发的模型进行模拟,并定量预测不均匀的细胞接种对扩增培养有不利影响,主要是通过降低增殖细胞的现存比例。所提出的模型有望用于预测成肌细胞行为,并设计这些细胞的高效扩增培养条件。