医学毕业人参二醇促骨髓cd34细胞增殖和分化的研究.docxVIP

XX大学 毕业论文 人参二醇促骨髓CD34+细胞增殖和分化的研究 姓 名： 2014年6月25日 作者：方桂伦高瑞兰林筱洁金锦梅 【摘要】 本研究探讨人参二醇(PDS)对正常人骨髓CD34+细胞的促进增殖 和诱导分化作用。用吸附单克隆抗体的免疫磁珠技术从正常人骨髓分离出CD34+ 细胞，采用琼脂半固体多向祖细胞集落(CFU Mix)培养方法观察PDS对CD34+ 细胞的增殖作用；用液体培养使CD344-细胞增殖，并向红系、粒系定向生长， 用流式细胞仪分析PDS诱导后细胞表而标记变化。结果显示：免疫磁珠分离 CD34+细胞的获得率占骨髓有核细胞(MNC)的(1.0±0.15) %；活细胞比例 占(95.6±1.2) %, CD34+ 细胞富集度达(86.8±2.8) %。中浓度 PDS (25 mg/L) 能够有效地促进CD34+细胞增殖，牛成CFU Mix集落，提高率为(56.3±3.5)%, 与对照相比较有显著差界(pO.Ol)。液体培养生成粒系、红系细胞的数量明显增 高，与对照相比差异有显著性，提高率分别为(35.6±3.2) %和(22.3±2.1) %, 与对照相比差异有显著性(pO.Ol),且呈剂量依赖性。经PDS诱导14天后，细胞 表面分化标记明显增高，CD33+细胞在PDS 50 mg/L时最高，CD71+细胞在 25 mg/L时最高，G A+细胞在10 mg/L时最高，而CD 15+细胞数量无明显变 化。结论：PDS不但促进CD34+细胞增殖，且能诱导其定向分化，提示PDS具 有类似造血生长因子和协同生长因子的效应。 【关键词】人参二醇 Effects of Ginseng Panaxadiol Saponin on Proliferation and Differentiation of Human Bone Marrow CD34+ Cells Abstract The study was purposed to investigate the effects of the panaxadiol saponin (PDS) from Ginseng on proliferation and differentiation of human CD34+ cells from human bone marrow. Highly purified CD34+ cells were isolated from human bone marrow by using the Dynal CD34 Cell Selection System (Dynal, Norway). The cells were exposed to PDS at various concentrations in both agar semi solid culture of CFU Mix and suspension culture of myeloid and erythroid cells in order to observe the efleets of PDS on proliferation of CD34+ cells. The cells were marked with 4 kinds of monoclonal antibody in related with their differentiation toward to myeloid and erythroid lineages, then examined by flow cytometry (FACS) after being incubated with PDS for 14 days. The results showed that the number of CD34+ cells was 1.0±0.15% out of marrow nuclear cells after being purified by Dynal beads system. The enrichment of CD34+ cells reached to 86.8±2.8 %. The best efficiency in promoting proliferation of CD34+ cells in vitro was obtained when the concentration of PDS was 25 mg/L, the formation of CFU Mix colonies significantly increased by 56.3±3.5% over those of no PDS control (pO.Ol)? The results from