脐血造血细胞扩增后细胞成分的变化

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。

Changes of cell component of amplified human umbilical blood hemapoietic cells

Objective To explore the optimal time for in vitro expansion of cord blood hemapoietic cells in quantity and quality.Methods Hemapoietic cells from human umbilical cord blood were cultured with rhIL-3,GM-CSF and rhEPO in a liquid system and colony forming cells and antigen expression were studied by colony forming assays and fluorescence-activated cell sorting.Results The number of cells increased at the 7th day and increased to the highest level at the 21st day by (8±4) fold and myeloid cells were in the majority.Lymphocytes (CD3 +、CD19 +) decreased at the 7th day and decreased to the lowest level after the 14th day.Percentage of CD34 + cells reached the highest level at the 14th day by (2.4±0.7)%,CD34 + cells increased by 30±7 fold,CD33 +CD34 + cell increased by 45±5 fold at the 21st day.CD71 + cells increased by (51±8)% at the 14th day,CD42a + cells increased to the highest level at the 21st day.The ability of cells to form colony was the highest at 7th day,but the colony-forming cells (CFCs) in total cultured cells were the highest at the 21st day.Conclusion The culture system containing the three growth factors expanded human umbilical cord blood hemapoietic cells:myeloid cell>erythroid cell>megakaryocyte;the optimal time for expansion was 21 days.