Mohme Malte, Maire Cecile L, Riecken Kristoffer, Zapf Svenja, Aranyossy Tim, Westphal Manfred, Lamszus Katrin, Fehse Boris
Laboratory for Brain Tumor Biology, Department of Neurosurgery, University Medical Center Hamburg-Eppendorf, 20246 Hamburg, Germany.
Research Department Cell and Gene Therapy, Department of Stem Cell Transplantation, University Medical Center Hamburg-Eppendorf, 20246 Hamburg, Germany.
Mol Ther. 2017 Mar 1;25(3):621-633. doi: 10.1016/j.ymthe.2016.12.014. Epub 2017 Jan 18.
Intratumoral heterogeneity has been identified as one of the strongest drivers of treatment resistance and tumor recurrence. Therefore, investigating the complex clonal architecture of tumors over time has become a major challenge in cancer research. We developed a new fluorescent "optical barcoding" technique that allows fast tracking, identification, and quantification of live cell clones in vitro and in vivo using flow cytometry (FC). We optically barcoded two cell lines derived from malignant glioma, an exemplary heterogeneous brain tumor. In agreement with mathematical combinatorics, we demonstrate that up to 41 clones can unambiguously be marked using six fluorescent proteins and a maximum of three colors per clone. We show that optical barcoding facilitates sensitive, precise, rapid, and inexpensive analysis of clonal composition kinetics of heterogeneous cell populations by FC. We further assessed the quantitative contribution of multiple clones to glioblastoma growth in vivo and we highlight the potential to recover individual viable cell clones by fluorescence-activated cell sorting. In summary, we demonstrate that optical barcoding is a powerful technique for clonal cell tracking in vitro and in vivo, rendering this approach a potent tool for studying the heterogeneity of complex tissues, in particular, cancer.
肿瘤内异质性已被确认为治疗耐药性和肿瘤复发的最强驱动因素之一。因此，随着时间推移研究肿瘤复杂的克隆结构已成为癌症研究中的一项重大挑战。我们开发了一种新的荧光“光学条形码”技术，该技术可使用流式细胞术（FC）在体外和体内对活细胞克隆进行快速追踪、识别和定量。我们对源自恶性胶质瘤（一种典型的异质性脑肿瘤）的两种细胞系进行了光学条形码标记。与数学组合学一致，我们证明使用六种荧光蛋白且每个克隆最多使用三种颜色，可明确标记多达41个克隆。我们表明光学条形码标记有助于通过流式细胞术对异质细胞群体的克隆组成动力学进行灵敏、精确、快速且廉价的分析。我们进一步评估了多个克隆对体内胶质母细胞瘤生长的定量贡献，并强调了通过荧光激活细胞分选回收单个活细胞克隆的潜力。总之，我们证明光学条形码标记是一种用于体外和体内克隆细胞追踪的强大技术，使该方法成为研究复杂组织（尤其是癌症）异质性的有力工具。
