定量PCR检测体外扩增脐带血造血干细胞HOXB4基因的表达  

Investigation of HOXB4 expression for ex vivo expanded cord blood stem cells (CBSCs)

作　　者：唐宇宏[1] 费小明[1] 沈文怡[1] 缪扣荣[1] 崔毓桂[2] 陆化[1] 汪承亚[1] 

机构地区：[1]南京医科大学第一附属医院血液科,江苏南京210029 [2]南京医科大学第一附属医院中心实验室,江苏南京210029

出　　处：《南京医科大学学报（自然科学版）》2005年第7期443-446,共4页Journal of Nanjing Medical University(Natural Sciences)

基　　金：江苏省卫生厅重点课题(H2001131);南京医科大学创新基金赞助项目(CX200104)

摘　　要：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 study the self-renewal potential of CBSCs cultured in vitro by detection of HOXB4 expression at mRNA level. Method: CBSCs were cultured for 7 or 14 days in serum-free media containing SCF, TPO, FL and G-CSF. Expression of HOXB4 at mRNA level was assayed by real-time RT-PCR. Results: The cell amount of CBSCs after culture increased greatly, however their HOXB4 expression declined compared with its pre-culture level. The average level of HOXB4 expression for 7-day-culture was around 18% of that of pre-cultured cells and the HOXB4 expression was as low as about 3% of its pre-culture level when they were cultured for 14 days. Conclusion: Results suggest that ex vivo expanded CBSCs will loss its self-renewal potential following culture in serum-free media gradually.

关 键 词：HOXB4 脐带血造血干细胞 体外扩增 实时定量RT-RCR 

分 类 号：R551[医药卫生—血液循环系统疾病]