The Primary Study of CD90(+)CD34(-)and Sca-1(+) Stem Cells Mobilized by EPO Plus G-CSF in Mice
Fang Zheng1, Long Cheng2, Qiang Yu2
1Institute of Hematology, Dongfeng Hospital affiliated to Yunyang Medical college, 16 Daling Avenue, Shiyan 442008, China. ; Center for Stem Cell Research and Application, Institute of Hematology, Union Hospital of Tongji Medical College, Huazhong University of Science and Technology, 1277 Jiefang Avenue, Wuhan 430022, China.
To investigate effects of recombinant human erythropoietin (rhEPO) plus recombinant human granulocyte colony stimulating factor (rhG-CSF) on mobilization of CD90(+)CD34(-) and Sca-1(+) stem cells in mice. rhEPO 1000 IU·kg(-1)·d(-1) per mice alone or plus rhG-CSF 250 μg·kg(-1)·d-1(-1)per mice was administered to mice for five days, the peripheral blood was collected at 6 hours, 1, 3,5 and 7 days after the last administration. The number of white blood cells (WBC) and mononuclear cells (MNC) was counted. The level of CD34, CD90 and Sca-1 mRNA were detected by reverse transcription-polymerase chain reaction (RT-PCR).The expressions of CD90(+)CD34(-) in absolute nuclear cells of peripheral blood was detected by flow cytometry. The results indicate that compared to mobilizing by rhG-CSF, rhEPO or plus rhG-CSF did not significantly enhance the number of WBC and MNC, on the fifth and the seventh day after the last administration, both the expression of CD34,CD90 and Sca-1 mRNA and the proportion of CD90(+)CD34(-) cells dramatically raised in rhEPO or in addition to rhG-CSF groups. We conclude that rhEPO or plus rhG-CSF had a strong capacity of mobilization of stem cells. The alteration of expression of CD34, CD90 and Sca-1 mRNA after the administration of rhEPO or combined with rhG-CSF indicated that this agents might potentially alter the peripheral blood graft content.
To investigate effects of recombinant human erythropoietin (rhEPO) plus recombinant human granulocyte colony stimulating factor (rhG-CSF) on mobilization of CD90(+)CD34(-) and Sca-1(+) stem cells in mice. rhEPO 1000 IU·kg(-1)·d(-1) per mice alone or plus rhG-CSF 250 μg·kg(-1)·d-1(-1)per mice was administered to mice for five days, the peripheral blood was collected at 6 hours, 1, 3,5 and 7 days after the last administration. The number of white blood cells (WBC) and mononuclear cells (MNC) was counted. The level of CD34, CD90 and Sca-1 mRNA were detected by reverse transcription-polymerase chain reaction (RT-PCR).The expressions of CD90(+)CD34(-) in absolute nuclear cells of peripheral blood was detected by flow cytometry. The results indicate that compared to mobilizing by rhG-CSF, rhEPO or plus rhG-CSF did not significantly enhance the number of WBC and MNC, on the fifth and the seventh day after the last administration, both the expression of CD34,CD90 and Sca-1 mRNA and the proportion of CD90(+)CD34(-) cells dramatically raised in rhEPO or in addition to rhG-CSF groups. We conclude that rhEPO or plus rhG-CSF had a strong capacity of mobilization of stem cells. The alteration of expression of CD34, CD90 and Sca-1 mRNA after the administration of rhEPO or combined with rhG-CSF indicated that this agents might potentially alter the peripheral blood graft content.
