人骨髓及脐血单个核细胞移植构建人源化非肥胖糖尿病/重症联合免疫缺陷小鼠模型:可以提高其人源化水平吗  

Does the level of human cell engraftment can be enhanced by constructing humanized nonobese diabetic/severe combined immunodeficient mouse models with transplantation of human bone marrow and cord blood mononuclear cell?

作　　者：赖颖晖[1] 赖永榕[1] 王明月[1] 杨高晖[1] 

机构地区：[1]广西医科大学第一附属医院血液科,广西壮族自治区南宁市530021

出　　处：《中国组织工程研究与临床康复》2009年第32期6249-6254,共6页Journal of Clinical Rehabilitative Tissue Engineering Research

基　　金：国家自然科学基金资助项目(30860307)~~

摘　　要：背景:人源化非肥胖糖尿病/重症联合免疫缺陷(nonobese diabetic/severe combined immunodeficient,NOD/SCID)小鼠模型在生物及医学科学研究中被广泛应用,而提高人源化NOD/SCID小鼠模型中的人源化水平是研究者的迫切需要。目的:探讨提高人源化NOD/SCID小鼠模型人源化水平的措施。设计、时间及地点:随机对照动物实验,于2008-11/2009-01在广西医科大学实验动物中心及广西医科大学第一附属医院完成。材料:雄性NOD/SCID小鼠14只,体质量(25.0±1.5)g;1例骨髓标本约6mL(多部位分层次采集)取自健康志愿者;1例足月脐带血标本大约90mL,取自广西医科大学第一附属医院产科,为健康产妇。方法:无菌抗凝的正常骨髓及足月脐带血样本均在采集后6h内处理。密度梯度离心法分离单个核细胞。将14只小鼠标号电脑随机分为5组:全身照射/环磷酰胺骨髓移植组(n=3),全身照射/环磷酰胺脐血移植组(n=3),全身照射骨髓移植组(n=3),全身照射脐血移植组(n=3),空白对照组(n=2)。将人骨髓或脐血单个核细胞分别移植给经不同方案和剂量预处理的各组小鼠,移植后给小鼠腹腔注射重组人粒-巨噬细胞集落刺激因子及重组人促红细胞生成素。主要观察指标:各组小鼠输注的细胞数,小鼠存活情况并计算死亡率,移植6周后流式细胞仪检测小鼠骨髓及外周血人CD45+细胞比例。结果:全身照射/环磷酰胺骨髓移植组、全身照射骨髓移植组和全身照射/环磷酰胺脐血移植组、全身照射脐血移植组小鼠输注人单个核细胞数分别为(0.6560±0.0089)×106和(4.0775±0.8450)×106。全身照射/环磷酰胺组死亡率均为100%,全身照射组死亡率均为33.3%。全身照射后再加环磷酰胺,小鼠不能耐受;而全身照射剂量越大,死亡率越高,当达400cGy时,死亡率100%。存活小鼠移植6周后,流式细胞仪检测小鼠骨髓中人CD45+细胞的比例,全身照射脐血移植组13号和6号小鼠�BACKGROUND： The humanized nonobese diabetic/severe combined immunodeficient （NOD/SCID） mouse models have been widely used in biological and medical research fields. Therefore, it is an urgent need to enhance the level of human cell engraftment in NOD/SCID mice models. OBJECTIVE： To investigate the measures that can enhance the level of human cell engraftment in the humanized NOD/SCID mice models. DESIGN, TIME AND SETTING： A randomized controlled animal experiment was performed at the experimental animal center and the First Affiliated Hospital of Guangxi Medical University between November 2008 and January 2009. MATERIALS： Fourteen male NOD/SCID mice, weighting （25.0＋1.5） g; a bone marrow prepare （approximate 6 mL） was harvested from a healthy volunteer; a full-term cord blood sample （approximate 90 mL） was provided by the Department of Obstetrics, First Affiliated Hospital of Guangxi Medical University. METHODS： The bone marrow and cord blood samples were treated within 6 hour after collection. And the mononuclear cells were isolated with density gradient centrifugation. Totally 14 mice were randomly divided into 5 groups. They were total-body irradiation/cyclophosphamide （TBI/CY） bone marrow transplantation group （n=3）, TBI/CY cord blood transplantation group （n=3）, TBI bone marrow transplantation group （n=3）, TBI cord blood transplantation group （n=3） and control group （n=2）. Each group mice were pretreated with the different schemes and dosage. Then human bone marrow or cord blood mononuclear cells were transplanted into mice by tail vein injection. After transplantation, human recombinant growth factors consisting of erythropoietin and granulocyte-macrophage colony-stimulating factor were injected into the mice intraperitoneally. MAIN OUTCOME MEASURES： The infused cell population, survival status of the mice, as well as was death rate calculated. At week 6 after the transplantation, the cells proportion of CD45＋ in mice bone marrow and peripheral blood

关 键 词：动物模型 NOD/SCID小鼠 人源化 细胞移植 

分 类 号：R394.2[医药卫生—医学遗传学]