健康人骨髓CD34^+造血干/祖细胞衰老与年龄的相关性研究  

Age-correlated studies of bone marrow CD34^+ hematopoietic stem/progenitor cells aging in healthy human

作　　者：耿珊 陈雄斌[2] 湛斌 封莎 季遥鹰 谢宝芬 GENG Shan;CHEN Xiong-bin;ZHAN Bin;FENG Sha;JI Yao-ying;XIE Bao-fen(Neurology Department,the 13th People＇Hospital of Chongqing,Chongqing 400053,China;Institute of Stem Cell and Tissue Engineering,Chongqing Medical University,Chongqing 400032,China)

机构地区：[1]重庆市第十三人民医院神内老年科,重庆400053 [2]重庆医科大学干细胞与组织工程实验室,重庆400032

出　　处：《局解手术学杂志》2018年第9期625-629,共5页Journal of Regional Anatomy and Operative Surgery

基　　金：重庆市卫生计生委医学科研计划项目(20143021)

摘　　要：目的探讨增龄过程中健康人CD34^+造血干/祖细胞(CD34^+HSC/HPCs)衰老的变化过程。方法由重庆医科大学附属第一医院血液科骨穿室提供正常人骨髓9例,将其分为青年组3例(女性31岁、男性26岁、男性28岁)、中年组3例(男性59岁、男性57岁、女性52岁)和老年组3例(男性70岁、男性70岁、女性62岁)。提取骨髓单个核细胞(BMNCs)后,免疫磁性细胞分选法分离纯化CD34^+细胞群,流式细胞术检测细胞纯度,台盼蓝染色检测细胞活性,衰老相关β-半乳糖苷酶(SA-β-gal)染色观察各年龄组阳性细胞百分比;造血干/祖细胞混合集落培养比较各年龄组CD34^+造血干/祖细胞形成混合集落能力。结果分选前每106个BMNCs中CD34^+细胞群比例为(1. 31±0. 54)%;免疫磁性分选后每106个细胞中CD34^+细胞群比例为(92. 5±0. 39)%。各年龄组人骨髓CD34^+造血干/祖细胞存活率可达99. 1%以上。人骨髓CD34^+造血干/祖细胞的SA-β-gal染色阳性率出现明显的随增龄变化趋势,老年组明显高于中年组及青年组。CD34^+造血干/祖细胞形成的干细胞混合集落能力随年龄增加降低。结论随年龄增加人骨髓CD34^+造血干/祖细胞出现明显衰老,表现为造血增殖能力下降,可能是造成一些老年人血液学疾病的原因。Objective To investigate the senescence process of CD34 ＋ hematopoietic stem/progenitor cells （CD34 ＋HSC/HPCs） in healthy human aging.Methods The human bone marrow without significant hematological abnormalities collected from 9 healthy people who were in hematology department of first affiliated hospital of Chongqing medical university were divided into three groups：youth group （3 cases：female 31 years old,male 26 years old,male 28 years old）,middle age group （3 cases：male 59 years,male 57 years,female 52 years old）,the elderly age group （3 cases：male 70 years old,male 70 years old,female 62 years old）.CD34 ＋ cells were separated and purified by immunomagnetic adsorption cell sorting （MACS） from BMNCs.Cell purity was detected by flow cytometry.Cell viability was assayed by trypan blue staining.Senescence-associated β-galactosidase（SA-β-gal） staining was used to observe the percentages of positive cells in all groups.Hematopoietic stem/progenitor cells mixed colony forming unit （CFU-Mix） culture was used to evaluate the colony forming ability of CD34 ＋ CD34 ＋HSC/HPCs of all groups.Results The proportion of CD34 ＋ cells per 10 6 BMNCs was （1.31± 0.54）% before sorting and （92.5±0.39）% per 10 6 cells after immunomagnetic sorting.The survival rate of CD34 ＋HSC/HPCs in human bone marrow was 99.1% in all groups.The positive rate of SA-β-gal staining of CD34 ＋HSC/HPCs in human bone marrow of elder group was significantly higher than that in middle-aged group or youth group.The number of CFU-Mix formed by CD34 ＋HSC/HPCs in young,middle and elderly patients decreased,and the number of cells in colonies decreased.Conclusion CD34 ＋ hematopoietic stem/ progenitor cells in human bone marrow show obviously senescent changes during aging,which may be the cause of some hematological diseases in the old human.

关 键 词：CD34+ 造血干/祖细胞 Β-半乳糖苷酶 混合集落能力 衰老 

分 类 号：R331.2[医药卫生—人体生理学]