Stimulation of bone marrow stromal cells conditioned medium on the expansion of mature megakaryocytes and colony forming unit-megakaryocyte in vitro
- Author:
- No author available
- Journal Title:
- ACTA PHYSIOLOGICA SINICA
- Issue:
- 2
- DOI:
- 10.3321/j.issn:0371-0874.2005.02.021
- Key Word:
- 骨髓;基质细胞;条件培养液;巨核细胞;扩增
Abstract： 为了研究骨髓成纤维细胞、内皮细胞和巨噬细胞条件培养液对巨核细胞的体外扩增作用,本文通过分离纯化骨髓成纤维细胞、内皮细胞和巨噬细胞,收集无血清骨髓成纤维细胞条件培养液(fibroblast conditioned medium,F-CM)、骨髓内皮细胞条件培养液(endothelial cell conditioned medium,E-CM)和骨髓巨噬细胞条件培养液(macrophage conditioned medium,M-CM),分别离心、超滤,获得原液、分子量(MW)＞10 kDa组分及MW＜10 kDa组分.将三种无血清条件培养液及其＞10 kDa组分和＜10 kDa组分,分别加入巨核细胞体外液体培养体系,观察成熟巨核细胞的生成.结果表明:F-CM和E-CM及其＞10 kDa组分加入培养体系后,成熟巨核细胞的生成显著增多,分别为对照组的(287.33±16.77)%、(141.21±17.47)%、(236.67±39.72)%和(179.03±30.98)%;F-CM的作用明显大于E-CM(P＜0.001);F-CM和E-CM的＜10 kDa组分、M-CM及其＞10 kDa组分和＜10 kDa组分加入培养体系后成熟巨核细胞数无明显改变.分别将F-CM、E-CM、M-CM加入巨核细胞液体扩增体系,24 h后取细胞作巨核细胞集落形成单位(colony forming unit-megakaryocyte,CFU-Meg)的测定.结果表明:F-CM和E-CM对CFU-Meg生成均有促进作用,M-CM对CFU-Meg生成无明显影响,扩增强度分别为扩增前的(168.18±30.24)%、(215.17±17.4)%和(85.0±7.0)%.逆转录聚合酶链式反应(reverse transcription-polymerase chain reaction,RT-PCR)实验结果表明:骨髓成纤维细胞表达促血小板生成素(thrombopoietin,TPO),不表达转化生长因子β1(transforming growth factor-β1,TGF-β1);骨髓内皮细胞表达TGF-β1,不表达TPO.上述结果提示:F-CM、E-CM及其＞10 kDa组分均对巨核系成熟细胞及巨核系祖细胞体外扩增有促进作用;F-CM对巨核系成熟细胞体外扩增的促进作用强于E-CM.
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