Isolation of Single Cells With the CellCelector for Subsequent Heterogeneity Analysis
Nearly all cellular systems are heterogeneous, contributing to specialized functionality and improved survival. A profound comprehension of single cell heterogeneity on genomic, epigenomic, transcriptomic and proteomic level is critical for understanding its impact on the functioning of organism in both healthy and diseased condition.
Most of our current knowledge about different cell and tissue types comes from bulk assays though, analyzing hundreds to millions of cells together, which may highly underestimate the true spectrum of cellular heterogeneity.
Modern technologies allow for single cell analysis, therefore taking a huge step forward in the understanding of cellular heterogeneity and its implications:
Single cell heterogeneity analysis can help to clarify developmental pathways, e.g. how stem cells make their fate decision, contributing to the development and maintenance of differentiated tissue. This is not only important for understanding malignancies caused by missing or misdirected differentiation but especially for the field of regenerative medicine.
Furthermore it is well known that tumor heterogeneity plays a crucial role in cancer evolution and clinical treatment, and is therefore a major contributor to therapy resistance and progression. A better understanding of tumors on single cell level unravelling tumor heterogeneity may have a huge impact on cancer survival rates. It is not only the heterogeneity of the tumor itself that influences the individual effectiveness of tumor therapy however but also mutations within the circulating tumor cells (CTCs). CTCs are tumor cells that have left the primary tumor to enter the blood stream and are driving metastasis.
In order to analyze the cellular heterogeneity on single cell level e.g. by whole genome amplification (WGA), single-nucleotide polymorphism (SNP) detection or sequencing target cells, they have to be selected and isolated in high purity from a heterogeneous background. The CellCelector is the perfect tool for the automated identification and isolation of 100% pure individual single cells offering many advantages over other methods.
Target cells are detected and selected for isolation based on microscopic imaging. This enables target cell selection utilizing fluorescent markers but also through distinct morphological features a target cell might express and that is visible in microscopy. This allows label free detection of target cells independently of fluorescent markers. Furthermore a live image during isolation as well as automated recording of before and after picking images of each isolated single cell create a comprehensive documentation of the picking process ensuring that only target cells have been picked without contamination of non-target cells. The fast isolation process is extremely gentle resulting in high cell integrity (up to 100% outgrowth rate in cloning applications) and outstanding transfer efficiency of up to 100%.
Related References
Segerman, A. et al. Clonal Variation in Drug and Radiation Response among Glioma-Initiating Cells Is Linked to Proneural-Mesenchymal Transition Cell Reports 17: 2994-3009 (2016)
- Neumann, M.H. et al. Isolation and characterization of circulation tumor cells using a novel workflow combining CellSearch® and CellCelector Biotechnol Prog. 33(1): 125-132 (2016)
- Yao, X. et al. Tumor cells are dislodged into the pulmonary vein during lobectomy J. Thorac Cardiovasc. Surg. 148(6): 3224-31 (2014)
- Heidary, M. et al. The dynamic range of circulating tumor DNA in metastatic breast cancer Breast Cancer Res. 16(4): 421 (2014)
- Lohr, G.J. et al. Whole-exome sequencing of circulating tumor cells provides a window into metastatic prostate cancer Nature Biotechnology 32: 479-484 (2014)