Mohseni Sofia, Khoshfetrat Ali Baradar, Rahbarghazi Reza, Khodabakhshaghdam Shahla, Kaleybar Leila Shafiei
Chemical Engineering Faculty, Sahand University of Technology, Tabriz, 51335-1996, Iran.
Stem Cell and Tissue Engineering Research Laboratory, Sahand University of Technology, Tabriz, 51335-1996, Iran.
J Biol Eng. 2023 Jun 5;17(1):38. doi: 10.1186/s13036-023-00358-4.
To evaluate shear stress influence on ex vivo expansion of hematopoietic cell lineages for clinical application, in this study, human pro-monocytic cell (namely U937 cell line) was selected as a hematopoietic stem cell (HSC) model and cultured in suspension mode at two different agitation rates (50, 100 rpm) in the stirred bioreactor. At the agitation rate of 50 rpm, the cells achieved higher expansion folds (27.4 fold) with minimal morphological changes as well as apoptotic cell death, while at 100 rpm the expansion fold decreased after 5-day of culture in suspension culture in comparison with static culture and reached 24.5 fold at the end of the culture. The results of glucose consumption and lactate production were also in agreement with the data of fold expansion and indicated the preference of culture in the stirred bioreactor when agitated at 50 rpm. This study indicated the stirred bioreactor system with an agitation rate of 50 rpm and surface aeration may be used as a potential dynamic culture system for clinical applications of hematopoietic cell lineage. The current experiments shed data related to the effect of shear stress on human U937 cells, as a hematopoietic cell model, to set a protocol for expansion of HSCs for biomedical applications.
为评估剪切应力对用于临床应用的造血细胞谱系体外扩增的影响,在本研究中,选择人原单核细胞(即U937细胞系)作为造血干细胞(HSC)模型,并在搅拌生物反应器中以两种不同的搅拌速率(50、100转/分钟)以悬浮模式进行培养。在50转/分钟的搅拌速率下,细胞实现了更高的扩增倍数(27.4倍),形态变化最小,凋亡细胞死亡也最少,而在100转/分钟时,与静态培养相比,悬浮培养5天后扩增倍数下降,培养结束时达到24.5倍。葡萄糖消耗和乳酸产生的结果也与扩增倍数数据一致,并表明在50转/分钟搅拌的搅拌生物反应器中培养更具优势。本研究表明,搅拌速率为50转/分钟且表面曝气的搅拌生物反应器系统可作为造血细胞谱系临床应用的潜在动态培养系统。当前实验提供了与剪切应力对作为造血细胞模型的人U937细胞的影响相关的数据,以制定用于生物医学应用的造血干细胞扩增方案。