Single-cell lineage tracing approaches in hematology research: technical considerations

• Published in: Experimental hematology Vol. 89; pp. 26 - 36
• Main Authors: Carrelha, Joana, Lin, Dawn S., Rodriguez-Fraticelli, Alejo E., Luis, Tiago C., Wilkinson, Adam C., Cabezas-Wallscheid, Nina, Tremblay, Cedric S., Haas, Simon
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier Inc 01.09.2020
• Subjects: Animals; Cell Differentiation; Cell Lineage - genetics; Cell Lineage - immunology; Cell Tracking - methods; Cell Transplantation - methods; Congresses as Topic; DNA Barcoding, Taxonomic - methods; Genetic Vectors - chemistry; Genetic Vectors - metabolism; Hematology - methods; Hematopoiesis - genetics; Hematopoiesis - immunology; Hematopoietic Stem Cells - cytology; Hematopoietic Stem Cells - immunology; Hematopoietic Stem Cells - virology; Homeostasis - genetics; Homeostasis - immunology; Humans; Lentivirus - genetics; Lentivirus - metabolism; Mice; Single-Cell Analysis - methods; Transgenes; Transposases - genetics; Transposases - immunology
• ISSN: 0301-472X; 1873-2399
• DOI: 10.1016/j.exphem.2020.07.007

•Clonal lineage tracing approaches for studying hematopoiesis are described.•Single-cell transplantation assays and lentiviral barcoding technologies are discussed.•Sleeping Beauty barcoding for studying cell fate in native hematopoiesis is covered. The coordinated differentiation of hematopoietic stem and progenitor cells (HSPCs) into the various mature blood cell types is responsible for sustaining blood and immune system homeostasis. The cell fate decisions underlying this important biological process are made at the level of single cells. Methods to trace the fate of single cells are therefore essential for understanding hematopoietic system activity in health and disease and have had a major impact on how we understand and represent hematopoiesis. Here, we discuss the basic methodologies and technical considerations for three important clonal assays: single-cell transplantation, lentiviral barcoding, and Sleeping Beauty barcoding. This perspective is a synthesis of presentations and discussions from the 2019 International Society for Experimental Hematology (ISEH) Annual Meeting New Investigator Technology Session and the 2019 ISEH Winter Webinar.