人胚胎AGM区基质细胞在体外培养中对脐血CD34^+细胞的支持作用  

Supportive effects of stromal cells from human embryonic aorta-gonad-mesonepheros region on umbilical cord blood CD34^+ cells

作　　者：陈惠芹[1] 张绪超[2] 黄绍良[2] 吴北燕[2] 吴燕峰[2] 蔡耘[2] 

机构地区：[1]中山大学附属第三医院儿科,广州510630 [2]中山大学附属第二医院干细胞研究中心,广州510120

出　　处：《中华血液学杂志》2006年第6期390-393,共4页Chinese Journal of Hematology

基　　金：国家自然科学基金(30300377);教育部博士点基金(20030558070);"十五"863计划资助项目(2003AA205008)

摘　　要：目的探讨人主功脉-性腺-中肾(AGM)区基质细胞对脐血 CD34^+细胞的造血支持作用。方法采用免疫磁珠法分离人脐血 CD34^+细胞,接种于底层已制备好的人 AGM 区基质细胞 S1～S5饲养层的24孔板中,同时设无饲养层组及取自同期胚胎的躯干成纤维(hFT)细胞为对照,体外培养28 d,每7 d 收获细胞并检测总数,流式细胞术检测 CD34^+、CD34^+ CD38^-细胞含量,并行造血细胞集落培养。结果在未加外源性造血生长因子的条件下,5株人 AGM 区基质细胞 hAGMS1～S5对制造血细胞总数、CD34^+及 CD34^+CD38^-细胞、造血细胞集落均有不同程度的扩增及维持作用,与无饲养层组、hFT 细胞组比较差异有统计学意义(P<0.05)。细胞总数在21d 达到峰值[扩增(25.13±4.83)倍],CD34^+、CD34^+CD38^-细胞在扩增14 d 达到峰值[(2.68±0.51)倍、(2.38±0.45)倍],高增殖潜能集落形成单位(HPP-CFU)亦在14 d 达到峰值[(2.62±0.85)倍]。hAGMS1～S5的造血支持作用组间比较差异亦有统计学意义(P<0.05),其中 hAGMS3、S4优于 S1、S2及 S5。结论人 AGM 区基质细胞对脐血 CD34^+细胞具有维持及扩增作用,特别是 hAGMS3、S4两株细胞具有更好的维持作用。Objective To explore the supportive effects of stromal cells from human aorta-gonadmesonephros（AGM） region on umbilical cord blood CD34^＋ cells. Methods Stromal cells derived from human AGM region （hAGMS1 -S5） and fetal trunk fibrobiasts （hFT） were cultured on the bottom of 24-well plates as feeder cells. CD34^＋ cells positively selected from human umbilical cord blood through immunomagnetic beads selection method, were seeded into 24-well plates, and eo-cuhured for 28d. The number of total nucleated cells （TNC） , CD34^＋ cells, CD34^＋ CD38^＋ cells,and CFC were counted every week. Results Stromal cells from human AGM region significantly supported proliferation of the TNC, CD3d^＋ cells, CD34^＋ CD38 cells and CFC, when compared with hFT and controls without feeder cells（P 〈0.05）. The TNC increased （25.13±4.83）- fold （peak value） at day 21 in group of co-culture with AGM stromal cells. CD34 ＋ and CD34 ＋ CD38^- cells increased （2.68± 0.51 ）- and （2.38 ± 0.45 ）- told respectively at day 14 of co-culture. In colony analysis, HPP-CFU increased （2.62 ± 0.85）- fold at day 14 of co-culture, The supportive effects of human AGM S1 -S5 were significantly different, hAGM S3 and S4 were better than hAGM S1 ,S2, and S5 （P 〈0.05）, Conclusions Human AGM stromal cells S1 -S5 could support the maintenance and expansion of umbilical cord blood CD34^＋ cells in vitro, hAGMS3, S4 call had better effects on maintaining HSC activity, whicb would provide model cells and basic data for researches on hematopoiesis mechanism and hematopoietic differentiation of embryonic stem cells.

关 键 词：主动脉-性腺-中肾区 基质细胞 造血干细胞 细胞培养 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]