Skip to main content
Fig. 3 | Stem Cell Research & Therapy

Fig. 3

From: miR-17 promotes expansion and adhesion of human cord blood CD34+ cells in vitro

Fig. 3

Effect of miR-17 modulation on the hematopoietic reconstitution potential of CB CD34+ cells in NPG mice. a Effect of miR-17 modulation on repopulation of CB CD34+ cells in NOD prkdcscid Il2rgnull (NPG™) mice. 4.0 × 104 miR-17 overexpression (17/OE), miR-17 knockdown (17/KD), or control (CTRL) CB CD34+ cells were injected intravenously into the sublethally irradiated NPG mice (n = 6 per group). The PB of NPG recipients was temporally monitored by analyzing CD45+ population every 4 weeks. The mice were sacrificed at 20 weeks after transplantation and the mononuclear cells from bone marrow were analyzed for human cells composed of CD45+, CD45CD36+ and CD36GPA+ cells and CD45+CD34+ population by flow cytometry. The level of total human cell engraftment is shown in the left panel. p < 0.03 or 0.04, between the NPG mice injected with 17/OE or 17/KD CD34+ cells and those injected with CTRL cells (Student’s t-test). The fraction of CD45+, erythroid (CD45CD36+ and CD36GPA+) and CD45+CD34+ population among the engrafted human cells is shown in the middle panel. The percent of CD45+ cells of PB from NPG recipients was shown in the right panel. *p < 0.05, between the NPG mice injected with 17/OE or 17/KD CD34+ cells and those injected with CTRL cells (Student’s t-test). The significant difference was analyzed between the mice injected with 17/OE or 17/KD CD34+ cells and those injected with CTRL cells. b Flow cytometry analysis of the human CB CD34+ cell repopulation in a representative NPG mouse after miR-17 modulation. Fresh CD34+ cells served as controls. The MNCs from bone marrow harvested from the engrafted NPG mice were examined by flow cytometry for the assessment of human cells composed of CD45+ cells (R1), erythroid cells including CD45CD36+ (R2) and CD36GPA+ (R3) populations and CD45+CD34+ cells (R4). c The levels of miR-17 expression in GFP-positive cells from engrafted mice at 20 weeks were tested by real-time PCR. d The bone marrow MNCs containing the different percentage of human cells (lanes 2–5) from the representative engrafted mice were analyzed for human-specific 17α-satellite DNA by PCR. The human-specific 17α-satellite gene was detected when the human cells were over 0.50 % (lanes 3–5) whereas it was indetectable at a percentage of 0.21 % (lane 2). Lane 1, one mouse without transplants; lane 2, one mouse receiving transplants of 17/OE CD34+ cells; lane 3, one mouse receiving transplants of 17/KD CD34+ cells; lane 4, one mouse receiving transplants of CTRL CD34+ cells; lane 5, one mouse receiving transplants of fresh CD34+ cells

Back to article page