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目的:通过检测HOXB4基因表达,评价体外扩增的脐带血干细胞(CBSC)自我更新的能力。方法:CBSC在含造血细胞因子(TPO+SCF+FL+G鄄CSF)的无血清培养基体外扩增7天和14天,抽提细胞总RNA,采用定量PCR检测HOXB4基因的表达。结果:CBSC体外培养后,细胞总数可以增加近100倍,但HOXB4的表达较扩增前下降。培养7天HOXB4的水平为扩增前的18%,培养14天,HOXB4表达仅为扩增前的3%左右。结论:CBSC体外扩增细胞数量增加的同时,自我更新能力逐渐下降,甚至消失。
OBJECTIVE: To evaluate the ability of expanded cord blood stem cells (CBSCs) to self-renew by detecting HOXB4 gene expression. Methods: CBSCs were expanded in serum-free medium containing hematopoietic cytokines (TPO + SCF + FL + G-CSF) for 7 days and 14 days. The total RNA was extracted and the expression of HOXB4 gene was detected by quantitative PCR. Results: After CBSC cultured in vitro, the total number of cells increased nearly 100-fold, but the expression of HOXB4 decreased compared with that before amplification. The level of HOXB4 cultured for 7 days was 18% before amplification. After cultured for 14 days, the expression of HOXB4 was only about 3% before amplification. Conclusion: While the number of CBSC expanded cells in vitro increases, the ability of self-renewal gradually decreases or even disappears.