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目的 探讨人脐血造血细胞体外扩增细胞数量和质量的变化及收获的最佳时机。方法 用重组人白细胞介素 - 3(rh IL- 3)、粒单集落刺激因子 (GM- CSF)和重组促红细胞生成素 (rh EPO)长期培养人脐血单个核细胞 ,动态观察其细胞构成和集落生成能力。结果 培养 1周后细胞总数逐渐增多 ,3周末细胞总数最多 ,扩增倍数为 (8± 4)倍 ,细胞构成则以髓系细胞为主 ;淋巴细胞 (CD3+ 、CD19+ )培养 1周明显下降 ,2周后低水平维持 ;培养 2周时 CD34+ 细胞百分比最高 ,为 (2 .4± 0 .7) % ,培养 3周时 CD34+ 细胞扩增倍数最大 ,为 (30± 7)倍 ,其中CD33+ CD34+ 细胞扩增倍数为 (4 5± 5 )倍。 CD71+ 细胞以培养 2周时最多 ,最高为 (5 1± 8) % ;CD42 a+ 细胞培养 3周时最高 ;单位数量细胞集落生成能力培养 ,1周时最大 ,但总的集落生成能力第 3周时最大。结论 含有上述生长因子的培养体系主要扩增脐血造血细胞的髓系细胞 ,次要扩增红系、巨核系细胞 ,最佳扩增时机为 3周。
Objective To investigate the changes of the number and mass of expanded human umbilical cord blood-derived cells in vitro and the optimal timing of harvesting. Methods Human umbilical cord blood mononuclear cells were cultured with recombinant human interleukin-3 (rhIL-3), granulocyte colony-stimulating factor (GM-CSF) and recombinant rhEPO (rh EPO) And colony-forming ability. Results The total number of cells increased gradually after 1 week of culture. The number of cells reached the maximum at 3 weeks, and the fold of expansion was (8 ± 4) times. The myeloid cells were the main component of cells. The number of lymphocytes (CD3 +, CD19 + After 2 weeks of culture, the percentage of CD34 + cells was the highest (2. 4 ± 0. 7)%, and the maximum fold of CD34 + cells was (30 ± 7) times at 3 weeks of culture. CD33 + CD34 + Cell multiplication was (4 5 ± 5) fold. The highest number of CD71 + cells was (51 ± 8)% at 2 weeks of culture. The highest number of CD42 a + cells at 3 weeks of culture was observed. The number of cells per colony forming ability was the highest at 1 week, but the total colony forming ability was the third week When the largest. CONCLUSION: The culture system containing the above-mentioned growth factors mainly expands the myeloid cells of cord blood hematopoietic cells. Secondary amplification of erythroid and megakaryocyte cells has the best timing of 3 weeks.