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The Influence of Co-culture Ex Vivo of CD34~+ Cells from Different Two Units Cord Blood on the Homing and Differentiation Ability of Each Other

Author: YaoZuo
Tutor: SunZiMin
School: Anhui Medical University,
Course: Internal Medicine
Keywords: Umbilical cord blood CD34 ~ cells In vitro co-culture Homing Differentiation
CLC: R329
Type: Master's thesis
Year: 2008
Downloads: 27
Quote: 0
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Abstract


Background and Purpose cord blood hematopoietic stem cells (haematopoietic stem cell, HSC) sources, cord blood contains the HSC a relatively small thus limiting its application in adults. Double mixed cord blood transplantation in the treatment of leukemia to be successful, to provoke their mechanism of interaction between. The study confirmed that patients receiving double umbilical cord blood transplantation, the majority of long-term hematopoietic only from a cord blood to a cord blood early in the implanted incurs exclusive, suggesting that there may be interactions between cord blood and cord blood. In this study, two cord blood CD34 - cells in vitro mixed culture homing and differentiation of each other by observing whether the impact of the understanding of the interaction of two cord blood is associated with the interaction between CD34 ~ cells. Amplification of mesenchymal stem cells (mesenchymal stem cells, MSC), isolated from normal human bone marrow the enrichment passage, the accelerator irradiation (12Gy) as trophoblast cells, immune beads election purified two cord blood CD34 ~ cells were inoculated to the surface (a CFSE-labeled) were incubated for 6 days the two CD34 to cell surface homing molecules detected by flow cytometry (CD26, CD44, CD54, CD62L, CD162, CD184) indicators of differentiation (CD33, CD41, CD71) changes. 1 human bone marrow MSC paste parietal Establishment and characterization of in vitro culture 12d, MSC fusion of 70% to 80%, was swirling growth. Cells spread to the third generation, the surface expression of CD90, CD105, CD166, CD29, CD44, CD13 and CD49e, did not express CD14, CD34, CD31, CD45 and HLA-DR. In vitro co-culture of each of the CD34 ~ cell homing ability to change cord blood CD34 ~ cell sorting enrichment purity (98.25 ± 0.93)%, the experimental group were incubated for 6 days, two cells of umbilical cord blood CD34 ~ proportion of reduced (60.4 ± 6.32)% and (60.2 ± 5.12)%, but no significant difference compared with the control group; experimental group of each of the CD34 ~ cells CD26, CD44, CD62L and CD184 positive rate than before culture There was no significant change. CD162 expression was significantly decreased. CD54 CD34 ~ cells ability to differentiate changes in the experimental group CD34 ~ cells were co-cultured for 3 days after the increase, but cultured for 6 days decreased to training level before, but no significant difference compared with the control group; 3 in vitro co-culture After incubation, the two CD34 ~ cells CD33 expression was decreased CD41 always low expression of CD71 marker expression increased, but there is still no significant difference compared with control group. The conclusion of the two cord blood CD34 ~ cells in vitro mixed culture homing and differentiation did not significantly affect each other, suggesting that the two cord blood interaction is not done through the interaction between CD34 ~ cells.

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