乙醛脱氢酶在检测脐带血造血干/祖细胞中的应用  

The application of acetaldehyde dehydrogenase in detection of cord blood haemopoietic stem/progenitor cells

作　　者：李莉 赵英新[2] 孟恒星[1] 刘兰婷 何祎 李茜[1,2] 韩俊领[1,2] 

机构地区：[1]协和干细胞基因工程有限公司,天津300384 [2]中国医学科学院北京协和医学院血液学研究所血液病医院,天津300020

出　　处：《生物医学工程与临床》2009年第4期324-327,共4页Biomedical Engineering and Clinical Medicine

摘　　要：目的通过比较乙醛脱氢酶(ALDH)与CD34、CD133及粒单核细胞集落形成单位(CFU-GM)在脐带血中含量的差异,探讨ALDH作为脐带血造血干/祖细胞(HSPC)检测指标的有用性。方法采集新鲜脐带血标本28份,用荧光底物法标记ALDH,流式细胞仪检测脐带血中低侧向角高表达ALDH活性(SSCloALDHbr)细胞、CD34+和CD133+含量,并进行CFU-GM的培养。结果脐带血SSCloALDHbr细胞表达率在ALDH反应后直接上机检测组(ALDH组)及在ALDH反应后进一步标记抗体再检测组(ALDH+抗体组)分别为(0.32±0.16)%和(0.30±0.17)%,两组相比差异无统计学意义(P>0.05)。脐带血CD34+表达率在抗体组和ALDH+抗体组分别为(0.40±0.26)%和(0.36±0.19)%,两组比较差异无统计学意义(P>0.05);脐带血CD133+表达率在抗体组和ALDH+抗体组分别为(0.18±0.16)%和(0.17±0.10)%,两组比较差异无统计学意义(P>0.05)。脐带血SSCloALDHbr细胞表达率和CD34+表达率、CD133+表达率及CFU-GM产率均呈正相关(r分别为0.87、0.69和0.54,P均<0.01)。结论ALDH反应后进一步标记抗体不影响脐带血SSCloALDHbr细胞的检测结果,ALDH反应亦不影响进一步标记抗体的检测结果;脐带血SSCloALDHbr细胞表达率和CD34+表达率、CD133+表达率及CFU-GM产率均呈正相关;ALDH活性检测可以作为检测脐带血HSPC的一个有用指标。Objective To study the usefulness of acetaldehyde dehydrogenase （ALDH） activity for characterizing cord blood haemopoietic stem/progenitor cell（HSPC） quality by comparison the differences of ALDH, CD34, CD 133, and the colony-forming units of granulocytes and monocytes（CFU-GM） in cord blood. Methods The samples （n = 28） of cord blood were harvested from placentas of full-term pregnancies immediately after delivery. The activity of ALDH from cord blood was detected by their fluorescent substrates. The expressions of side-scatter low ALDH brigbt（SSC^loALDH^br cells, CD34^＋ cells, and CD 133^＋ cells were measured by fluorescence- activated cell sorting（FACS）. The CFU-GM were also evaluated by the colony-forming units（CFU） assay. Results The expressions of SSC^loALDH^br cells in the direct analysis group after ALDH reaction （ALDH group） and the further labeled antibody group （ALDH ＋ antibody group） were （0.32 ± 0.16） % and（0.30 ± 0.17） %, respectively. There was no significant difference between 2 groups（P〉 0.05）. The expressions of CD34^＋ were （0.40 ± 0.26） % in the direct labeling antibody group（antibody group） and （0.36 ± 0.19） % in the ALDH ＋ antibody group, respectively. There was no significant difference between 2 groups （P〉 0.05）. Moreover, the expressions of CD 133^＋ were （0.18 ± 0.16） % in antibody group and （0.17 ± 0.10） % in the ALDH ＋ antibody group, respectively. There was also no significant difference between 2 groups （P 〉 0.05 ）. The activity of ALDH in cord blood was positive correlation with the expressions of CD34^＋, CD133^＋ and the yield of CFU-GM （r = 0.87, 0.69, 0.54, P 〈 0.01）. Conclusion The detection results of SSC^loALDH^br cells in cord blood could not impacted by further labeling antibody after ALDH reaction, and the detection results of further labeled antibody also could not impacted by ALDH reaction. The activity of ALDH is positive correlation with the expressions of CD34^＋,

关 键 词：脐带血 干/祖细胞 SSC^loALDH^br细胞 CD34^+ CD133^+ 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学]