摘要
目的:观察人胎盘造血干细胞对非致死辐射小鼠移植模型造血功能重建的作用。方法:采用机械处理联合酶解法提取人胎盘来源造血干细胞(PHSC),利用流式细胞术和细胞集落形成实验进行胎盘HSC鉴定。选取25只NOG小鼠随机分为空白对照组(n=5),辐照组(n=4),低剂量PHSC组(n=8)和高剂量PHSC组(n=8)。在无菌条件下对辐射组、低剂量PHSC组和高剂量PHSC组小鼠分别进行X-射线照射,每只1 Gy(100cGy/min)。给予低剂量PHSC组和高剂量PHSC组小鼠尾静脉注射0.1 ml人胎盘HSC,数量分别为2×10^6/ml和1×10^7/ml,空白对照组和辐射组小鼠给予等体积生理盐水注射。每周对小鼠进行称重并计算相对体重变化,分别于第4、8和12周抽取各组小鼠外周血进行人CD45+和髓系、淋巴系血细胞流式细胞术检测。结果:流式细胞术检测和细胞集落培养鉴定显示,人胎盘HSC占人胎盘单个核细胞的比例达到0.75%以上且具有造血干细胞的分化能力。与空白对照组小鼠比较,辐照组、低剂量PHSC组和高剂量PHSC组小鼠相对体重增长幅度显著下降;与辐照组小鼠比较,低剂量PHSC组和高剂量PHSC组小鼠相对体重增长幅度显著升高。流式细胞术结果显示,在移植后12周,高剂量PHSC组小鼠中人血液免疫系统开始长期重建,而低剂量PHSC小鼠人CD45+细胞的比例非常低。结论:将人胎盘HSC移植入非致死辐射小鼠体内,能够获得人血细胞短期重建和长期重建,证明人胎盘HSC能够在辐射小鼠体内多向分化并重建造血功能。
Objective:To investigate the effect of human placental hematopoietic stem cells(PHSCs)on hematopoietic reconstruction in non-lethally irradiated mice.Methods:Human placental HSCs were extracted by mechanical method combined with zymolysis and were identified by flow cytometry and colony formationtest.Twenty-five NOG mice were divided randomly into 4 groups:the blank control group(n=5),the irradiated group(n=4),the low dose PHSC group(n=8)and the high dose PHSC group(n=8).The mice in the irradiated,the low dose and the high dose PHSC groups were irradiated with X-rays at dose 1 Gy(100 cGy/min)under sterile condition.The mice in the low dose PHSC group and the high dose PHSC group were injected intravenously with 0.1 ml human placental HSC in dose of 2×10^6 and1×10^7,respectively.The mice in the blank control and the irradiated group were injected with the same volume of saline.The mice were weighed weekly,and the changes of body weight were calculated.The peripheral blood was collected from each group at 4,8 and 12 week for flow cytometrytic detection of human CD45+and myeloid and lymphoid cells.Results:The flow cytometry and cell-colong formation test showed that the human placental HSC accounted for more than 0.75%of total placental mononuclear cells,moreover possess the differentiation ability.Compared with the blank control group,the relative weight gain in the irradiated,the low dose PHSC,and the high dose PHSC groups decreased significantly,and the relative weight gain in the low dose PHSC and the high dose PHSC group increased significantly as compared with the irradiated group.Flow cytometry showed that at the tine-point of 12 weeks after transplantation,the human blood immune system in the high-dose PHSC mice began long-term reconstruction,while the ratio of human CD45+cells in the low-dose PHSC mice was very low.Conclusion:After transplantation of human PHSC the nonlethally irradiated mice can obtain short-term and long-term reconstruetion of human blood cells,which demonstrated that human placental HSC can differentiate and reconstruct hematopoietic function in vivo of irradiated mice.
作者
季夏芸
朱丰城
李天宇
周新
王浩
JI Xia-Yun;ZHU Feng-Cheng;LI Tian-Yu;ZHOU Xin;WANG Hao(Cruilife Stem Cell Science and Technology Co.,Ltd,Wuxi 214023,Jiangsu Province,China;Department of Hematology,Wuxi People's Hospital/Wuxi Children's Hospital Affiliated to Nanjing Medical University,Wuxi 214023,Jiangsu Province,China;Department of Hematology,Wuxi People's Hospital Affiliated to Nanjing Medical University,Wuxi 214023,Jiangsu Province,China)
出处
《中国实验血液学杂志》
CAS
CSCD
北大核心
2019年第6期1986-1992,共7页
Journal of Experimental Hematology
基金
无锡市科技发展资金(CBE01G1603)
江苏省科教强卫创新团队(CXTDB2017016)
无锡市科教强卫发展学科(FZXK001)
无锡市卫生局重大项目(Z201302)
关键词
人胎盘造血干细胞
辐射小鼠
造血功能
human placental hematopoietic stem cells
irradiated mice
hematopoietic function
