Involvement of the PI3K/AKT Pathway in the Formation and Fusion of Spheroids Derived from Human Dermal Fibroblast for Tissue Engineering Technology
Involvement of the PI3K/AKT Pathway in the Formation and Fusion of Spheroids Derived from Human Dermal Fibroblast for Tissue Engineering Technology
复制标题
DOI：
10.1134/s1990519x22040022
复制
发表时间：
2022-08-01
影响因子：
--
通讯作者：
Kamohara, K.
中科院分区：
文献类型：
--
作者：
Amamoto, S.;Itoh, M.;Kamohara, K.
通过利用细胞聚集体(球体)的生物制造技术，成功地从人真皮成纤维细胞的自体细胞中制造出了人造血管，并有望应用于临床。但是，生产周期仍然是一个主要问题，必须优化组织结构。人真皮成纤维细胞(HDFB)来源的多细胞球体(MCS)的形成和融合是组织构建的基础，因此有必要研究影响MCS形成和融合的因素。在本研究中，我们评估了HDFB衍生的MCS的形成和融合作为制造过程的基础，重点是使用MCS的生物制造技术中的PI3K/AKT级联。用HDFB评价MCS的形成和融合程度。转化生长因子-β-1作为PI3K/AKT通路的激活剂，而LY294002和CoA-Cl作为其抑制剂。与对照组相比，抑制PI3K/AKT通路显著抑制MCS的形成和融合，而激活PI3K/AKT通路显著促进MCS的形成和融合。我们在PI3K/AKT途径中发现了6个与细胞黏附相关的基因(ITGB1、PTK2、MAP2K1、PDK1、GJA1和PI3KR1)，并证实了基因表达的显著差异。本研究表明，抑制PI3K/AKT级联可抑制骨髓间充质干细胞的形成和融合，而转化生长因子-β-1激活该级联通路可促进骨髓间充质干细胞的形成和融合。本研究有助于阐明骨髓间充质干细胞的形成和融合机制，优化组织工程技术基础，促进再生医学的发展。
Through the biofabrication technology using cellular aggregates (spheroids), an artificial blood vessel derived from autologous cells of human dermal fibroblast was successfully produced and is expected to be applied clinically. However, the production period remains a major problem, and the organizational construction must be optimized. It is necessary to focus on the formation and fusion of multicellular spheroids (MCS) derived from human dermal fibroblast (HDFB) and consider what affects MCS formation and fusion because these are the basis of tissue construction. In this study, we evaluated HDFB-derived MCS formation and fusion as the basis of the manufacturing process, focusing on the PI3K/AKT cascade in biofabrication technology using MCS. HDFB were used to assess the degree of MCS formation and fusion. TGF-β1was used as an activator of the PI3K/AKT pathway, whereas LY294002 and COA-Cl were used as its inhibitors. Compared with the control group, the suppression of the PI3K/AKT pathway significantly inhibited MCS formation and fusion, whereas the activation of the PI3K/AKT pathway significantly promoted MCS formation and fusion. We identified six genes (ITGB1,PTK2,MAP2K1,PDK1,GJA1, andPI3KR1) related to cell adhesion in the PI3K/AKT pathways and confirmed a significant difference in gene expression. This study showed that the formation and fusion of HDFB-derived MCS were inhibited by suppressing the PI3K/AKT cascade, whereas it was promoted by activating the cascade with TGF-β1. This research may help elucidate the mechanism of MCS formation and fusion and optimize MCS and tissue construction, which are the basis for tissue engineering technology, and contribute to the development of regenerative medicine.