Traject3d allows label-free identification of distinct co-occurring phenotypes within 3D culture by live imaging.
Traject3d allows label-free identification of distinct co-occurring phenotypes within 3D culture by live imaging.
复制标题
轨迹3D允许通过实时成像在3D培养中无标记鉴定3D培养物中不同的同时发生表型。
DOI：
10.1038/s41467-022-32958-x
复制
发表时间：
2022-09-09
影响因子：
16.6
通讯作者：
中科院分区：
文献类型：
--
作者：
通过遗传学、蛋白质组学和成像方法进行的单细胞分析扩大了识别调节不同细胞状态的程序的能力。细胞或组织碎片的三维（3D）培养提供了一个系统来研究这些状态如何有助于多细胞形态发生。由于缺乏检测这种异质性的工具，接种到3D培养物中的细胞是否产生单一表型或是否平行产生多种生物学上不同的表型在很大程度上是未知的。在这里，我们开发了Traject3d（3D中的轨迹识别），这是一种用于识别3D培养中异质状态的方法，以及这些状态如何随着时间的推移从无标记的多日延时成像中产生不同的表型。我们用它来描述癌细胞系形态状态的时间景观，在转移潜力和耐药性方面各不相同，并使用这些信息来识别抑制这种异质性的药物组合。因此，Traject3d是其他单细胞技术的重要伴侣，通过实时成像促进实时识别不同状态如何导致在3D培养中平行发生的替代表型。目前缺乏检测3D培养物异质性的工具。在这里，作者报告了Traject3d作为一个框架来识别3D文化中的异质状态，并了解这些状态如何使用无标记的多日延时成像产生不同的表型。
Single cell profiling by genetic, proteomic and imaging methods has expanded the ability to identify programmes regulating distinct cell states. The 3-dimensional (3D) culture of cells or tissue fragments provides a system to study how such states contribute to multicellular morphogenesis. Whether cells plated into 3D cultures give rise to a singular phenotype or whether multiple biologically distinct phenotypes arise in parallel is largely unknown due to a lack of tools to detect such heterogeneity. Here we develop Traject3d (Trajectory identification in 3D), a method for identifying heterogeneous states in 3D culture and how these give rise to distinct phenotypes over time, from label-free multi-day time-lapse imaging. We use this to characterise the temporal landscape of morphological states of cancer cell lines, varying in metastatic potential and drug resistance, and use this information to identify drug combinations that inhibit such heterogeneity. Traject3d is therefore an important companion to other single-cell technologies by facilitating real-time identification via live imaging of how distinct states can lead to alternate phenotypes that occur in parallel in 3D culture. There are currently a lack of tools to detect heterogeneity in 3D cultures. Here the authors report Traject3d as a framework to identify heterogeneous states in 3D culture and to understand how these give rise to distinct phenotypes using label-free multi-day time-lapse imaging.
登录
查看更多内容
影响因子：
48
作者：
Huber W;Carey VJ;Gentleman R;Anders S;Carlson M;Carvalho BS;Bravo HC;Davis S;Gatto L;Girke T;Gottardo R;Hahne F;Hansen KD;Irizarry RA;Lawrence M;Love MI;MacDonald J;Obenchain V;Oleś AK;Pagès H;Reyes A;Shannon P;Smyth GK;Tenenbaum D;Waldron L;Morgan M
通讯作者：
Morgan M
影响因子：
8
作者：
Horiguchi, K.;Sakamoto, K.;Koinuma, D.;Semba, K.;Inoue, A.;Inoue, S.;Fujii, H.;Yamaguchi, A.;Miyazawa, K.;Miyazono, K.;Saitoh, M.
通讯作者：
Saitoh, M.
影响因子：
4.3
作者：
Chen H;Lau MC;Wong MT;Newell EW;Poidinger M;Chen J
通讯作者：
Chen J
影响因子：
12.3
作者：
Carpenter AE;Jones TR;Lamprecht MR;Clarke C;Kang IH;Friman O;Guertin DA;Chang JH;Lindquist RA;Moffat J;Golland P;Sabatini DM
通讯作者：
Sabatini DM
影响因子：
8
作者：
Esquer H;Zhou Q;Nemkov T;Abraham AD;Rinaldetti S;Chen YC;Zhang X;Orman MV;D'Alessandro A;Ferrer M;Messersmith WA;LaBarbera DV
通讯作者：
LaBarbera DV