小鼠脾集落形成期基质细胞在体外扩增脐带血CD34^＋细胞中的作用

目的：研究来源于小鼠脾脏的基质细胞在体外扩增脐血CD34^ 细胞中的作用;方法：取γ射线照射后小鼠脾集落形成期的脾细胞经人重组单核细胞集落刺激因子（rhM-CSF)的激发培养,获得贴壁脾基质细胞,经丝裂霉素处理后作为滋养细胞,与多种造血细胞生长因子共同应用,体外扩增经免疫磁珠阳性选择法纯化的人脐血CD34^ 细胞,用体外集落形成及流式细胞仪分析扩增细胞的集落形成能力和表型。结果：1）rhM-CSF能有效地刺激小鼠脾基质细胞的生长;2）所获的小鼠脾基质细胞具有支持脐血CD34^ 细胞的集落形成能力,与多种造血细胞生长因子共同应用后,能有效地扩增CD34^ 细胞,扩增2周后的细胞数达100倍,且仍具有多向分化能力。结论：rhM－CSF能刺激小鼠脾集落形成期基质细胞的增殖,该类基质细胞具有有效支持人脐血CD34^ 细胞的体外扩增作用。

Effects of recombinant human macrophage colony stimulating factor (rhM-CSF) on stromal cell derived during the time of mice colony formation unit-spleen (CFU-S) and its role on CD34+ cells expansion in vitro

Objective To investigate the effects of stromal cell derived from mice splee n during in vitro ex pansion of CD34 hematopoietic stem cells from umbilical cord blood.Methods Irradiated and non-irradiated mice at day10,12and14were sacrificed and their spleens wer e taken for collecting spl -een cells.The cells were stimulated with recombinan t human macrophage colony stimulating factor(rhM-CSF)for collecting adherent spleen derived stroma cells.The effects of spleen derived stromal cells which treated with mitomycin served as a feeder layer,combined with different cytok ines on in vitro exp ansion of purified cord blood CD34 were assayed.Clonogenic assay and flow cytometry were emp -loyed to analyze phenotype of expanded cells and their ability to form different colony in vitr o.Re s-ults1)rhM-CSF efficiently stimulated spleen derived stromal cells to p roliferate;2)The mice str-omal cells obtained during the time of colony form ation-unit spleen(CFU-S)promoted the CD34 cells to form colony in semisolid medium and,combined with cytokines,there by efficiently expanded the hematopoietic cells from cord blood.After two week s incubation,CD34positive cells were100times more than original cells cult ured and expanded cells still have the ability to form mu lti-lineage col-ony in vitro.Conclusion rhM-CSF can stimulate the stromal cells derived from CFU-S to proli fe-rate and support the survival and expansion of CD34 cells separated from cord blood.