Figure 3. Altered mobilization ability in diabetic mice is not cell autonomous but microenviroment dependent.

(A) Schematic representation of the procedure followed to assess whether the inhibition of mobilization is cell autonomous or microenvironment dependent. Lethally irradiated SJL recipients were transplanted with 1×10⁶ BM cells from STZ-treated or control mice, along with 1×10⁶ CD45.1 support cells. 16 weeks after transplantation the SJL recipients from the two groups underwent G-CSF mobilization regimen. (B) Number of CFU-C in the PB of recipients transplanted with STZ-treated or control cells after the induction of mobilization with G-CSF. Columns represent mean ± s.e.m., n=8. (C) Actual numbers and (D) ratio of CD45.1 and CD45.2 positive cells in the peripheral blood of recipient mice transplanted with STZ-treated or control cells before and after mobilization with G-CSF treatment. Columns represent mean ± s.e.m., n=9 and 8 respectively. (E) Schematic representation of the reverse experiment in which 15 STZ-treated and control C57Bl6 mice were transplanted with wild type CD45.1 whole bone marrow cells. After 16-weeks 10 mice were evaluated for LSK and LT-HSC numbers. 5 mice underwent G-CSF mobilization therapy followed by blood collection and evaluation of CFU-C number. (F) Graph bars represent number of LKS per femur. Data are mean ± s.e.m., n=10, ** p<0.01. (G) Number per femur of LT-HSC Lin⁻Sca⁺cKit⁺CD150⁺CD48⁻ and (H) Lin- Lin⁻Sca⁺cKit⁺Flk2⁻CD34⁻ as assessed by flow cytometry. Data are mean ± s.e.m., n=10. (I) Number of CFU-C per 100 μl of G-CSF-mobilized peripheral blood from STZ-treated and control mice 16 weeks after transplantation, data are mean ± s.e.m., n=5, * p<0.05.