Project description:Background: Circulating tumor cells (CTCs) are cells that have broken off from a primary solid tumor, entered the blood stream, and have the potential to metastasize to nearby tissues. Thus, CTCs represent an important biomarker to monitor cancer progression and patient outcome. The isolation, identification and characterization of CTCs are critical for this goal. We previously developed a method for the detection and isolation of melanoma CTCs from peripheral blood through negative and positive antibody enrichment. Methods: Here we show that this protocol is easily applied to other cancers; in this case we expand and validate this method to include prostate cancer. Results: We show that this combined negative and positive enrichment protocol successfully isolates CTCs for characterization using single cell RNA sequencing and have updated the protocol to include a newer version of single cell library preparation, Smart-seq3. In our analysis, cells prepared for sequencing with Smart-seq3, which uses unique molecular identifiers (UMIs), showed no considerable difference compared to those prepared using the previous iteration of the protocol, Smart-seq2. Conclusion: This study demonstrates a robust and flexible method for the isolation and characterization of CTCs from various cancers for potential use as biomarkers for disease management and for characterization with single cell RNA-seq.
Project description:The olfactory mucosa (OM) has the unique characteristic of pursuing almost continuous neurogeneration during adulthood in response to external injuries to maintain olfactory function due to the presence of stem cells. The easily accessibility of the OM in humans is a feature that makes these stem cells relevant candidates for the development of regenerative therapies. In this report we present an in depth characterization of patient derived OM together with the characterization of their respective OM derived cultures. In addition, we present a method for the enrichment and isolation of OM stem cells that can be used for future translational sought studies regarding plasticity, neuro-regeneration or disease modeling.
