Tomita Shunsuke, Sakao Miho, Kurita Ryoji, Niwa Osamu, Yoshimoto Keitaro
Biomedical Research Institute , National Institute of Advanced Industrial Science and Technology , 1-1-1 Higashi , Tsukuba , Ibaraki 305-8566 , Japan . Email:
College of Arts and Sciences , The University of Tokyo , 3-8-1 Komaba , Meguro , Tokyo 153-8902 , Japan . Email:
Chem Sci. 2015 Oct 1;6(10):5831-5836. doi: 10.1039/c5sc01259g. Epub 2015 Jun 30.
Currently available methods for stem cell evaluation require both prior knowledge of specific markers and invasive cell lysis or staining, hampering the development of stem cell products with assured safety and quality. Here, we present a strategy using optical cross-reactive sensor arrays for markerless and noninvasive identification of differentiated stem cell lineages with common laboratory equipment. The sensor array consists of a library of polyion complexes (PICs) between anionic enzymes and synthetic poly(ethylene glycol)-modified polyamines, which can recognize "secretomic signatures" in cell culture supernatants. Due to the reversible nature of PIC formation, the incubation of diluted culture supernatants with PICs caused enzyme release through competitive interactions between the secreted molecules and the PICs, generating unique patterns of recovery in enzyme activity for individual cell types or lineages. Linear discriminant analysis of the patterns allowed not only normal/cancer cell discrimination but also lineage identification of osteogenic and adipogenic differentiation of human mesenchymal stem cells, therefore providing an effective way to characterize cultured cells in the fields of regenerative medicine, tissue engineering and cell biology.
目前可用的干细胞评估方法既需要特定标志物的先验知识,又需要进行侵入性细胞裂解或染色,这阻碍了具有可靠安全性和质量的干细胞产品的开发。在此,我们提出一种策略,利用光学交叉反应传感器阵列,借助普通实验室设备对分化的干细胞谱系进行无标记和非侵入性识别。该传感器阵列由阴离子酶与合成聚(乙二醇)修饰的多胺之间的聚离子复合物(PICs)库组成,可识别细胞培养上清液中的“分泌组特征”。由于PIC形成的可逆性,将稀释的培养上清液与PICs孵育会通过分泌分子与PICs之间的竞争性相互作用导致酶释放,从而为单个细胞类型或谱系产生独特的酶活性恢复模式。对这些模式进行线性判别分析不仅可以区分正常/癌细胞,还可以鉴定人间充质干细胞成骨和成脂分化的谱系,因此为再生医学、组织工程和细胞生物学领域中培养细胞的表征提供了一种有效方法。