title: Arrayed lentiviral barcoding for quantification analysis of hematopoietic dynamics.
abstract: Our understanding of system dynamics of mixed cell populations in whole organisms has benefited from the advent of individual cell marking by nonarrayed DNA barcodes subsequently analyzed by high-throughput DNA sequencing. However, key limitations include statistical biases compromising quantification and the lack of applicability to deconvolute individual cell fate in vivo after pooling single cells differentially exposed to different conditions ex vivo. Here, we have derived an arrayed lentiviral library of DNA barcodes and obtained a proof-of-concept of its resolving capacity by quantifying hematopoietic regeneration after engraftment of mice with genetically modified autologous cells. This method has helped clarify and bridge the seemingly opposed clonal-succession and continuous-recruitment models of hematopoietic stem cell behavior and revealed that myeloid-lymphoid biases are common occurrences in steady-state hematopoiesis. Arrayed lentiviral barcoding should prove a versatile and powerful approach to deconvolute cell dynamics in vivo with applications in hematology, embryology, and cancer biology.
authors.affiliation: CEA, Institute of Emerging Diseases and Innovative Therapies (iMETI), F-92265 Fontenay-aux-Roses, Paris, France; Inserm U962 and University Paris Sud 11, CEA-iMETI, F-92265 Fontenay-aux-Roses, Paris, France.
authors.forename: Jeanne
authors.initials: J
authors.lastname: Grosselin
authors.forename: Karine
authors.initials: K
authors.lastname: Sii-Felice
authors.forename: Emmanuel
authors.initials: E
authors.lastname: Payen
authors.forename: Stany
authors.initials: S
authors.lastname: Chretien
authors.forename: Diana
authors.initials: D
authors.lastname: Tronik-Le Roux
authors.forename: Philippe
authors.initials: P
authors.lastname: Leboulch
mesh_terms: D000818:Animals; D058948:Cell Tracking / Q000379:methods; D058893:DNA Barcoding, Taxonomic; D005822:Genetic Vectors; D057809:HEK293 Cells; D018380:Hematopoietic Stem Cell Transplantation; D006412:Hematopoietic Stem Cells / Q000502:physiology*; D006801:Humans; D016086:Lentivirus / Q000235:genetics*; D051379:Mice; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D010641:Phenotype
keywords: Hematopoiesis; Hematopoietic stem cells; In vivo tracking; Lentiviral vector