【摘要】 目的:检测小鼠原始造血细胞c-kit基因表达以评价体外模拟骨髓造血微环境。方法:建立微载体-基质细胞体外造血模型。根据已发表的小鼠c-kit基因序列设计引物,采用RT-PCR方法检测c-kit mRNA水平,并测定祖细胞集落产率。结果:小鼠骨髓造血细胞2周培养实验显示,粒系-巨噬系造血祖细胞集落产率(CFU-GM/10~5):模型组比液体悬浮培养和单纯微载体基质细胞培养对照组集落产率之总和高2.1倍(t=2.869,P<0.05);原始造血细胞的c-kit基因表达水平(OD比率):模型组比两个对照组之总和高3.1倍(t=2.858,P<0.05)。结论:在没有外加细胞因子的条件下,微载体-基质细胞造血模型可抑制造血干、祖细胞过度分化与耗竭,维持其c-kit mRNA较高的表达水平。更多还原

【Abstract】 Objective: To evaluate ex vivo hematopoietic microenviroment of stimulating bone marrow by detecting c-kit gene expression of primary hematopoietic cells. Methods: The hematopoietic model for stromal cell adhering to microcarrier was established. A pair of oligonucleotide primer from the published mouse c-kit DNA sequence were used in RT-PCR to test the level of c-kit mRNA, and the colony yield of hematopoietic progenitor cells were determined. Results: We set up the model group (G1) in comparison with only microarrier-stromal cell culture group (G2) and only marrow cell culture group (G3). The experiments of hematopoietic cell culture for two weeks showed: (1) the yield of colony forming unit of granulocyte-macrophage (CFU-GM/105): Gl was 2.1-fold as much as the sum of G2 and G3 yield (t = 2.869,P<0.05); (2) c-kit gene expression level of primary hematopoietic cells (OD ratio) : G1 was 3.1-fold as high as the sum of two control groups levels (t =2.858,P<0.05). Conclusion: Under the condition of supplementing without cytokines, the hematopoietic model can inhibit hematopoietic stem and progenitor cells from excessively differentiating and exhausting, and maintain the higher level of expressing c-kit mRNA.更多还原