title: Analysis of the clonal growth and differentiation dynamics of primitive barcoded human cord blood cells in NSG mice.
abstract: Human cord blood (CB) offers an attractive source of cells for clinical transplants because of its rich content of cells with sustained repopulating ability in spite of an apparent deficiency of cells with rapid reconstituting ability. Nevertheless, the clonal dynamics of nonlimiting CB transplants remain poorly understood. To begin to address this question, we exposed CD34+ CB cells to a library of barcoded lentiviruses and used massively parallel sequencing to quantify the clonal distributions of lymphoid and myeloid cells subsequently detected in sequential marrow aspirates obtained from 2 primary NOD/SCID-IL2Rγ(-/-) mice, each transplanted with ∼10(5) of these cells, and for another 6 months in 2 secondary recipients. Of the 196 clones identified, 68 were detected at 4 weeks posttransplant and were often lympho-myeloid. The rest were detected later, after variable periods up to 13 months posttransplant, but with generally increasing stability throughout time, and they included clones in which different lineages were detected. However, definitive evidence of individual cells capable of generating T-, B-, and myeloid cells, for over a year, and self-renewal of this potential was also obtained. These findings highlight the caveats and utility of this model to analyze human hematopoietic stem cell control in vivo.
authors.affiliation: Terry Fox Laboratory, BC Cancer Agency, Vancouver, BC, Canada;
authors.forename: Alice M S
authors.initials: AM
authors.lastname: Cheung
authors.forename: Long V
authors.initials: LV
authors.lastname: Nguyen
authors.forename: Annaick
authors.initials: A
authors.lastname: Carles
authors.forename: Philip
authors.initials: P
authors.lastname: Beer
authors.forename: Paul H
authors.initials: PH
authors.lastname: Miller
authors.forename: David J H F
authors.initials: DJ
authors.lastname: Knapp
authors.forename: Kiran
authors.initials: K
authors.lastname: Dhillon
authors.forename: Martin
authors.initials: M
authors.lastname: Hirst
authors.forename: Connie J
authors.initials: CJ
authors.lastname: Eaves
mesh_terms: D000818:Animals; D018952:Antigens, CD34 / Q000378:metabolism; D001483:Base Sequence; D002454:Cell Differentiation; D019070:Cell Lineage; D049109:Cell Proliferation; D002999:Clone Cells / Q000145:classification / Q000166:cytology / Q000378:metabolism; D036101:Cord Blood Stem Cell Transplantation / Q000379:methods*; D058893:DNA Barcoding, Taxonomic; D005312:Fetal Blood / Q000166:cytology* / Q000378:metabolism; D005434:Flow Cytometry; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D006367:HeLa Cells; D006801:Humans; D016130:Immunophenotyping; D053631:Interleukin Receptor Common gamma Subunit / Q000172:deficiency / Q000235:genetics; D007700:Kinetics; D051379:Mice; D016688:Mice, Inbred NOD; D018345:Mice, Knockout; D016513:Mice, SCID; D008969:Molecular Sequence Data; D009841:Oligonucleotides / Q000235:genetics; D013997:Time Factors; D014183:Transplantation, Heterologous
chemical_list: D018952:Antigens, CD34; C507942:Il2rg protein, mouse; D053631:Interleukin Receptor Common gamma Subunit; D009841:Oligonucleotides; D049452:Green Fluorescent Proteins
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