摘要
目的 比较骨髓与脐带血细胞体外培养基质细胞的基本特性 ,为基质细胞的选择应用提供依据。方法 用Dexter长期培养体系培养骨髓和脐带血基质细胞 ,以细胞增殖、细胞形态、细胞化学染色、细胞表面及基质细胞支持另一骨髓细胞形成的鹅卵石造血区 (CAFC) ,长期培养起始细胞 (LTC IC)为指标 ,比较两者的生长特性、成分及功能。结果 ①细胞生长特性 :出现贴壁细胞时间 ,骨髓细胞为培养 3d ,脐带血细胞为培养 5～ 6d ;细胞融合成片时间 ,骨髓细胞为培养 10～ 14d ,脐带血细胞为培养 12～ 18d ;第 2 1天细胞增殖数 ,骨髓比脐带血细胞增殖少 ;②细胞成分 :2 1d培养后细胞成分 ,骨髓来源者以成纤维细胞为主 ,其次是巨噬细胞与内皮细胞 ,脂肪细胞最少 ;脐带血细胞来源者 ,以巨噬细胞为主 ,其次是内皮细胞、成纤维细胞 ,偶见脂肪细胞 ;细胞化学与上述结果基本一致 ;细胞表面抗原检测 ,CD14、CD4 5的表达骨髓细胞明显低于脐带血细胞 ;③细胞功能 :骨髓来源的基质细胞较脐带血细胞的基质细胞支持另一骨髓细胞形成的CAFC和LTC IC明显多。结论 ①生长特征 :形成贴壁细胞时间骨髓较脐带血短 ,骨髓细胞比脐带血细胞有核细胞数增殖快、持续时间相对短 ;②细胞成分特性 :骨髓来源形成的基质细胞以成纤维细胞为主 。
Objective Compare the basic feature of the stromal cells derived from bone marrow and cord blood culture in vitro,and provide laboratory data for its clinical usage.Methods Dexter long term culture system was used for bone marrow and cord blood's stromal cells culture.Cell proliferation was evaluated by quantity and morphology study of the cells.Components of the stromal cells were detected by histochemistry staining assay and surface antigen expression analysis.Cobble stone area forming cells (CAFC) and long term culture initiating cells (LTC IC) assay was employed to evaluate the function of the stromal cells.Results Adherent cells appeared after 3 days culture from bone marrow and 5～6 days from cord blood.Cells confluence was detected at 10～14 days in bone marrow culture and 12～18 days in cord blood culture.The proliferation of nucleated cells in bone marrow culture was less significant than that in cord blood culture at 21 days.Cells components assay at 21 days suggested that fibroblasts were predominant in bone marrow culture,then were macrophages and endothelial cells,adipocytes were the fewest.While in cord blood culture,macrophages were predominant,the quantity of endothelial cells and macrophages were smaller,and adipocytes were hardly observed.CAFC and LTC IC form ability of stromal cells derived from bone marrow was stronger than that derived from cord blood.Conclusions ①The time for bone marrow culture to forming adherent cells was less than that for cord blood culture.The proliferation of nucleated cells in bone marrow culture was earlier than that in cord blood culture,which maintained longer time proliferation.②Fibroblasts were predominant in stromal cells from bone marrow culture,while macrophages were predominant in cord blood culture.③stromal cells derived from bone marrow supported better for CAFC and LTC IC growing than that from cord blood.
出处
《北京医学》
CAS
北大核心
2002年第6期399-402,共4页
Beijing Medical Journal
