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目的 探讨人骨髓基质细胞 (hBMSC)协同以干细胞因子和FL为主的细胞因子对脐血CD3 4 + 细胞的体外扩增作用。方法 采用免疫磁珠法分选脐血CD3 4 + 细胞 ,以SCF +IL 3+IL 6 +FL +EPO组合高效扩增CD3 4 +细胞[1] ,并结合该细胞因子组合接种到预先照射 (2 0Gy)的hBMSC上 ,d10结束培养 ,收获细胞分别作细胞计数、集落培养和流式细胞术检测CD3 4 + 细胞数。结果 本法获得的脐血CD3 4 + 细胞纯度较高 (92± 0 .0 4 ) % ,在hBMSC组培养的d2 ,造血细胞几乎都粘附到hBMSC上 ,随着培养时间的延长 ,CD3 4 + 细胞比例不断下降。hBMSC组与无hBMSC组相比 ,除细胞总数扩增倍数外 ,CFU GM、BFU E、CD3 4 + 细胞扩增倍数差异有显著性意义 (P <0 .0 5 )。结论 ①脐血来源的CD3 4 + 细胞粘附于滋养层上形成造血灶 ,且 10d后造血细胞仍具有体外集落形成能力 ,表明骨髓基质细胞可支持并维系体外造血 ;②hBMSC协同外源性细胞因子可能是扩增造血干 /祖细胞的较理想方案
Objective To investigate the in vitro expansion of cord blood CD3 4 + cells by human bone marrow stromal cells (hBMSC) in combination with cytokines such as stem cell factor and FL. Methods Cord blood CD3 4 + cells were sorted by immunomagnetic beads and CD3 4 + cells were expanded efficiently by SCF + IL 3 + IL 6 + FL + EPO combination [1]. The CD3 + 2 0Gy) hBMSC, d10 end culture, harvested cells were counted as cells, colony culture and flow cytometry CD3 4 + cells. Results The purity of cord blood CD3 4 + cells obtained by this method was higher than that of hBMSCs group (92 ± 0. 04%). In the hBMSC group, almost all the hematopoietic cells adhered to the hBMSCs. With the prolongation of culture time, CD3 4 + The proportion of cells is declining. Compared with non-hBMSC group, hBMSC group had significant difference in multiplication of CFU GM, BFU E and CD34 + cells except multiplication of cell number (P <0.05). Conclusion ① CD3 4 + cells derived from umbilical cord blood adhere to the trophoblast and form hematopoietic foci. After 10 days, hematopoietic cells still have the colony forming ability in vitro, indicating that bone marrow stromal cells can support and maintain hematopoiesis in vitro. ② hBMSCs cooperate with exogenous cytokines It may be a more ideal method to amplify hematopoietic stem / progenitor cells