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探讨胎儿、儿童、成人骨髓基质细胞对脐血造血细胞体外长期生存的维持能力 ,以及细胞因子组合在基质接触和无基质的培养体系中扩增脐血干、祖细胞的效应 .将连续二次Ficoll Hypaque分离的脐血单个核细胞(MNCs)接种至经辐照预处理的各种骨髓基质细胞上 ,持续培养 6周 ,定期检测祖细胞 (CFC)和长期培养起始细胞 (LTC IC) ;使用干细胞因子 (SCF)、白细胞介素 (IL) 3、IL 6组合 ,在基质接触和无基质体系中 ,扩增脐血干、祖细胞 .结果表明 :①成人骨髓基质细胞能更有效地维持脐血 2周、4周CFCs的数量 ,在 10 5初始接种MNCs中 ,产量分别为 6 4 5± 11 3和 43 6± 7 6 ;而胎儿骨髓基质细胞对脐血LTC ICs表现出更强的支持作用 ,其LTC ICs的产量为 18 5± 5 2 .②成人基质细胞接触的扩增体系 ,CFCs扩增最为显著 ,其CFCs的产量为扩增前的 8 4倍 ;胎儿基质接触的扩增体系能够更有效地扩增LTC ICs ,其产量为扩增前的 3 9倍 .基于上述结果 ,认为不同发育阶段的骨髓基质细胞对脐血CFCs和LTC ICs的生存 ,表现出不同的维持效应 ;基质接触的扩增体系更适合于脐血原始造血细胞的扩增 .
To investigate the ability of fetal, pediatric and adult bone marrow stromal cells to maintain the long-term survival of cord blood hematopoietic cells in vitro and the effects of cytokine combination on cord blood stem and progenitor cells in matrix-contacted and stroma-free culture systems. Cord blood mononuclear cells (MNCs) isolated from Ficoll Hypaque were seeded on irradiated bone marrow stromal cells for 6 weeks, and the progenitor cells (CFCs) and long-term culture initiating cells (LTC ICs) were detected regularly. Umbilical cord blood stem / progenitor cells were expanded in matrix-contacted and matrix-free systems using combinations of stem cell factor (SCF), interleukin (IL) 3, and IL 6. The results showed that: ① adult bone marrow stromal cells can be more effectively maintained The numbers of CFCs in cord blood at 2 and 4 weeks of cord blood were 6 4 5 ± 11 3 and 43 6 ± 7 6, respectively, in 10 5 initial inoculation MNCs, whereas fetal marrow stromal cells showed stronger cord blood LTC ICs The production of LTC ICs was 18 5 ± 5 2. (2) The proliferation of adult stromal cells exposed to CFCs was the most significant, with CFCs yielding 84% of that before amplification. The fetal stromal contact amplification The system can more efficiently amplify LT Based on the above results, it is considered that bone marrow stromal cells in different developmental stages have different maintenance effects on the survival of cord blood CFCs and LTC ICs. The matrix contact amplification system is more Suitable for cord blood hematopoietic cell expansion.