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目的双份脐血移植的植入动力学机制目前尚无定论,推测双份脐血中的淋巴细胞与优势份脐血的产生相关,本实验将双份脐血的CD34+细胞与CD3+细胞混合培养,观察CD3+细胞对CD34+细胞的增殖分化有无影响。方法建立液体和半固体培养体系,将免疫磁珠分选纯化的双份脐血间的CD34+细胞和CD3+细胞混合培养6d和14d。以流式细胞计数观测CD34+细胞培养后的分化指标(CD33,CD41,CD71);计数集落形成单位(GM-CFU、BFU-E、GEMM-CFU)分析CD34+细胞的增殖情况。结果液体共培养后各份CD34+细胞表面分化指标的变化。脐血CD34+细胞分选富集的纯度为(98.70±0.72)%。3d实验组和对照组的各项分化指标无差异(P>0.05);6d的CD33、CD71实验组明显低于对照组,而CD41明显高于对照组(P<0.05)。半固体共培养后CD34+细胞增殖能力的变化。实验组的红系集落形成单位(BFU-E)及粒单细胞集落形成单位(GM-CFU)数低于对照组(P<0.05),而混合细胞集落形成数(GEMM-CFU)高于对照组(P<0.05)。结论将两份脐血的CD34+细胞和CD3+细胞体外混合培养对CD34+细胞的增殖分化能力有影响,推测双份脐血间的相互作用可部分地通过CD3+细胞介导。
OBJECTIVE: To study the implanted kinetic mechanism of double cord blood transplantation is inconclusive. It is speculated that the lymphocytes in double cord blood are associated with the production of dominant cord blood. In this experiment, double cord blood CD34 + cells were mixed with CD3 + cells To observe whether CD3 + cells have any effect on the proliferation and differentiation of CD34 + cells. Methods The liquid and semi-solid culture system was established. CD34 + cells and CD3 + cells in double-labeled cord blood were separated and purified by magnetic beads for 6 days and 14 days. Differentiation index (CD33, CD41, CD71) of CD34 + cells were observed by flow cytometry. Proliferation of CD34 + cells was analyzed by counting colony forming units (GM-CFU, BFU-E and GEMM-CFU). Results The changes of the surface differentiation of CD34 + cells after liquid co-culture. The purity of cord blood CD34 + cells was (98.70 ± 0.72)%. There was no significant difference in the differentiation index between the 3d experimental group and the control group (P> 0.05). On the 6th day, the CD33 and CD71 experimental groups were significantly lower than the control group, while the CD41 was significantly higher than the control group (P <0.05). Proliferation of CD34 + Cells after Semi - solid Coculture. The number of BFU-E and GM-CFU in the experimental group was lower than that in the control group (P <0.05), while the number of mixed colony-forming units (GEMM-CFU) Group (P <0.05). Conclusions The in vitro mixed culture of CD34 + cells and CD3 + cells from two cord blood cells has an impact on the ability of CD34 + cells to proliferate and differentiate. It is speculated that the interaction between double copies of umbilical cord blood can be partly mediated by CD3 + cells.