Deep imaging of bone marrow shows non-dividing stem cells …

Posted: August 23, 2016 at 12:42 am

a, Hindlimb bone marrow cellularity (n=9 mice for -catulin+/+,n=4 mice for -catulinGFP/+ and n=9 mice for -catulinGFP/GFP genotype) and spleen cellularity (n=6 mice for -catulin+/+,n=4 mice for -catulinGFP/+ and n=6 mice for -catulinGFP/GFP genotype), spleen mass (7 mice for -catulin+/+,n=4 mice for -catulinGFP/+ and n=7 mice for -catulinGFP/GFP genotype). b, White blood cell (WBC), red blood cell (RBC) and platelet (PLT) counts per microliter of peripheral blood from 812 week old -catulin+/+, -catulinGFP/+, and -catulinGFP/GFP mice (n=9 mice/genotype). c,d, Frequencies of mature hematopoietic cells and progenitors in the bone marrow of 812 week old -catulin+/+ and -catulinGFP/GFP mice (Pre-ProB cells were B220+sIgMCD43+CD24; ProB cells were B220+sIgMCD43+CD24+; Pre-B cells were B220+sIgMCD43; common lymphoid progenitors (CLPs) were Linc-kitlowSca1lowCD127+CD135+; common myeloid progenitors (CMPs) were Linc-kit+Sca1CD34+CD16/32; granulocyte-macrophage progenitors (GMPs) were Linc-kit+Sca1CD34+CD16/32+; and megakaryocyte-erythroid progenitors (MEPs) were Linc-kit+Sca1CD34CD16/32 (n=3 mice/genotype). e, Bone marrow CD150+CD48LSK HSC frequency, bone marrow CD150CD48LSK MPP frequency (n=12 mice/genotype in 12 independent experiments), and spleen HSC frequency (n=3 mice/genotype in 3 experiments). f, Percentage of HSCs and whole bone marrow cells that incorporated a 3 day pulse of BrdU in vivo (n=6 -catulin+/+, 9 -catulinGFP/+, and 7 -catulinGFP/GFP 812 week old mice in 3 independent experiments). g, Colony formation by HSCs in methylcellulose cultures (GM means granulocyte-macrophage colonies, GEMM means granulocyte-erythroid-macrophage-megakaryocyte colonies, Mk means megakaryocyte colonies; (n=5 mice/genotype in 5 independent experiments). h, Reconstitution of irradiated mice by 300,000 donor bone marrow cells from 812 week old -catulin+/+, -catulinGFP/+, or -catulinGFP/GFP mice competed against 300,000 recipient bone marrow cells (n=4 donor mice and 16 recipient mice for -catulin+/+, n=3 donor mice and 9 recipient mice for -catulinGFP/+, and n=4 donor mice and 18 recipients for -catulinGFP/GFP in 3 independent experiments). i, Serial transplantation of 3 million WBM cells from primary recipient mice shown in panel d into irradiated secondary recipient mice (n=4 primary -catulin+/+ recipients were transplanted into 17 secondary recipients and n=6 primary -catulinGFP/GFP recipients were transplanted into 20 secondary recipients). All data represent means.d. The statistical significance of differences between genotypes was assessed using Students t-tests or ANOVAs. None were significant.

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