造血調控因子HAPO、PF4對造血干細胞調節(jié)的體內外研究
本文關鍵詞:造血調控因子HAPO、PF4對造血干細胞調節(jié)的體內外研究 出處:《中國協(xié)和醫(yī)科大學》2005年博士論文 論文類型:學位論文
更多相關文章: 血液血管細胞生成素 CD133~+細胞 增殖 粘附性 趨化作用 造血重建 化療 減毒沙門氏菌 PF4 5-氟尿嘧啶
【摘要】:目的 研究人促血液血管細胞生成素(Hemangiopoietin,HAPO)對CD133~+造血干細胞的作用,探討其生物學功能。方法 采用細胞液體培養(yǎng)、半固體培養(yǎng)、免疫熒光標記流式細胞儀測定、顯微鏡觀察照相等方法。結果 1.在加SCF、TPO、IL-3的無血清無基質培養(yǎng)體系中,培養(yǎng)的人骨髓CD133~+細胞,,7天時流式細胞儀測定對照組CD133~+細胞含量為41.92%,HAPO組CD133~+細胞含量為56.70%;HAPO組中CD133~+細胞數(shù)由原來的6.8×10~4增加為(2.88±0.12)×10~6,細胞數(shù)擴增了41.98倍;對照組中CD133~+細胞數(shù)由原來的6.8×10~4增加為(1.79±0.02)×10~6,細胞數(shù)擴增了26.09倍;10天時流式細胞儀測定對照組CD133~+細胞含量為32.60%,HAPO組CD133~+細胞含量為49.02%;HAPO組中CD133~+細胞數(shù)擴增了356倍,而對照組中CD133~+細胞數(shù)擴增了98倍。2.擴增7天時,流式細胞儀測定對照組細胞粘附分子CD31細胞含量對照組為68.27%,HAPO組CD31細胞含量為76.89%;細胞粘附分子CD49d表達對照組80.11%、HAPO組CD49d細胞含量為88.17%。3.將5×10~3個CD133~+細胞接種到1ml甲基纖維素半固體集落培養(yǎng)基中,PBS對照組生成GM-CFU 134.5±9.2個,HAPO組生成GM-CFU 167.5±4.5個,
[Abstract]:Objective to study the effect of human Hemangiopoietin (HAPO) on CD133~+ hematopoietic stem cells and to explore its biological function. Methods the methods of cell culture, semisolid culture, immunofluorescent labeling flow cytometer and microscope observation were used. Results 1. in SCF, TPO, IL-3 and serum-free medium culture system, human bone marrow CD133~+ cells cultured for 7 days, when the flow cytometry content of CD133~+ cells control group 41.92%, HAPO group of CD133~+ cells was 56.70%; the number of CD133~+ cells in the HAPO group from the original 4 increased to 6.8 * 10~ (2.88 + 0.12) * 10~6, the cell number was 41.98 times; the control group: the number of CD133~+ cells from 6.8 * 10~4 to increase (1.79 + 0.02) * 10~6, the cell number was 26.09 times; 10 days were determined by flow cytometry in CD133~+ cells of control group was 32.60%, HAPO group of CD133~+ cells the content is 49.02%; the number of CD133~+ cells in HAPO group was 356 times, while the control group in the number of CD133~+ cells was 98 times. 2. after 7 days of amplification, flow cytometry was used to determine the cell adhesion molecule CD31 cell content in the control group, 68.27% in the control group, and 76.89% in the HAPO group, while the CD49d expression in the cell adhesion molecule was 80.11% in the control group and 88.17% in the HAPO group, and the CD49d cell content in the CD31 group was 88.17%. 3., 5 x 10~3 CD133~+ cells were inoculated into 1ml semi-solid medium, and PBS control group generated GM-CFU 134.5 + 9.2, HAPO group generated GM-CFU 167.5 + 4.5.
【學位授予單位】:中國協(xié)和醫(yī)科大學
【學位級別】:博士
【學位授予年份】:2005
【分類號】:R329
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