Experimental design and clonal characteristics of hematopoiesis. (A) Schematic diagram of the rhesus macaque autologous transplantation model. CD34⁺ cells were collected following mobilization and transduced with a lentiviral vector containing a 6 base-pair library ID followed by a highly diverse 27 or 35 base-pair randomly generated barcode library. After total body irradiation, the transduced CD34⁺ cells were reinfused back into the autologous macaque, and T cells (T), B cells (B), monocytes (Mo), and granulocytes (Gr) were purified after transplantation. Barcode retrieval was performed by polymerase chain reaction of genomic DNA and illumina sequencing. (B) Transplantation characteristics of 2 aged (RQ3600, RQ859) and 3 young (ZH33, ZG66, ZK22) macaques. (C) Percentage of GFP⁺ cells in hematopoietic lineages across time after transplantation. (D) Cumulative number of independent barcoded clones detected above the threshold at a minimum of 1 time point is shown for each hematopoietic lineage and the total for all lineages across time. Clone numbers were calculated after applying a threshold of a clone achieving a fractional read abundance of at least 0.05% in at least 1 cell type at a minimum of at least 1 time point. The plateaus indicate highly sensitive capture of contributing clones. (E) Number of unique barcoded clones detected at each time point in each hematopoietic lineage and in all lineages. (F) Shannon diversity of each lineage and for all lineages. The Shannon diversity index encompasses both the number of clones and the evenness of their distribution.