Hirono Keita, Hayashi Yusuke, Ogawa Yuuki, Kino-Oka Masahiro, Sugiyama Hirokazu
Department of Chemical System Engineering, The University of Tokyo, Tokyo, Japan.
Department of Biotechnology, Graduate School of Engineering, The University of Osaka, Osaka, Japan.
Biotechnol Bioeng. 2025 Apr 25. doi: 10.1002/bit.29001.
With their many therapeutic functions, mesenchymal stem cells (MSCs) are promising sources for regenerative medicine. However, in the manufacture of MSCs, without a method for exploring the effects of long-term passage on cell proliferation potentials, the design of passage culture processes is challenging. Here, for the process design of the MSC passage culture, we propose a model for predicting the growth rate as a function of the cumulative population doubling level (cPDL) for each passage. Three steps were implemented: (1) passage culture experiments to correlate apparent growth rate with cPDL were conducted, (2) a model for predicting the growth rate as a function of cPDL was developed, and (3) a model to design the passage culture of MSCs from bone marrow (BM-MSCs) and umbilical cord (UC-MSCs) with stochastic simulation was applied. Two design variables (passage number and harvesting time) were investigated to define feasible operation regions as probabilistic design spaces to meet three quality indicators (senescence level, confluency level, and total number of cells) with given probabilities. Consequently, 10 and 62 conditions out of 165 were identified as feasible for BM- and UC-MSCs, respectively, which would contribute to the industrial MSC passage culture process design.
间充质干细胞（MSCs）具有多种治疗功能，是再生医学中很有前景的细胞来源。然而，在MSCs的生产过程中，由于缺乏探索长期传代对细胞增殖潜能影响的方法，传代培养过程的设计具有挑战性。在此，为了进行MSCs传代培养的过程设计，我们提出了一个模型，用于预测每次传代时生长速率作为累积群体倍增水平（cPDL）的函数。该模型的建立分为三个步骤：（1）进行传代培养实验，以关联表观生长速率与cPDL；（2）建立一个预测生长速率作为cPDL函数的模型；（3）应用一个通过随机模拟设计骨髓间充质干细胞（BM-MSCs）和脐带间充质干细胞（UC-MSCs）传代培养的模型。研究了两个设计变量（传代次数和收获时间），以确定可行的操作区域作为概率设计空间，从而以给定概率满足三个质量指标（衰老水平、汇合度水平和细胞总数）。结果，在165种条件中，分别确定了10种和62种条件对BM-MSCs和UC-MSCs是可行的，这将有助于工业上MSCs传代培养过程的设计。
