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中华移植杂志(电子版)

中华移植杂志(电子版) ›› 2019, Vol. 13 ›› Issue (04) : 298 -301. doi: 10.3877/cma.j.issn.1674-3903.2019.04.010

所属专题： 文献；

论著

第三方脐血细胞辅助输注对小鼠单倍型造血干细胞移植后造血重建的影响
杜振兰1, 陈鹏2,†()   
  1. 1. 100007　北京，解放军总医院第七医学中心附属八一儿童医院血液和肿瘤科　出生缺陷防控关键技术国家工程实验室　儿童器官功能衰竭北京市重点实验室
    2. 100007　北京，解放军总医院第七医学中心血液科
  • 收稿日期:2019-04-09 出版日期:2019-11-25
  • 通信作者: 陈鹏

The influence of umbilical cord blood cells infusion on hematopoietic reconstitution in a haploidentical hematopoietic stem cell transplantation mouse model

Zhenlan Du1, Peng Chen2,†()   

  1. 1. Department of Heamotology and Oncology, Affiliated BAYI Children′s Hospital, the Seventh Medical Center of PLA General Hospital, National Engineering Laboratory for Birth Defects Prevention and Control of Key Technology, Beijng Key Laboratory of Pediatric Organ Failure, Beijing 100007, China
    2. Department of Heamotology, the Seventh Medical Center of PLA General Hospital, Beijing 100007, China
  • Received:2019-04-09 Published:2019-11-25
  • Corresponding author: Peng Chen
  • About author:
    Corresponding author: Chen Peng, Email:
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杜振兰, 陈鹏. 第三方脐血细胞辅助输注对小鼠单倍型造血干细胞移植后造血重建的影响[J]. 中华移植杂志(电子版), 2019, 13(04): 298-301.

Zhenlan Du, Peng Chen. The influence of umbilical cord blood cells infusion on hematopoietic reconstitution in a haploidentical hematopoietic stem cell transplantation mouse model[J]. Chinese Journal of Transplantation(Electronic Edition), 2019, 13(04): 298-301.

目的

分析以脐血细胞作为第三方细胞辅助输注对小鼠单倍型造血干细胞移植后造血重建的影响。

方法

以CB6/F1雄性小鼠为供鼠，以BALB/C小鼠为受鼠。受鼠经Co60全身照射清髓后回输供鼠干细胞，建立单倍型造血干细胞移植模型。将40只受鼠分为对照组和实验组，每组各20只，对照组受鼠回输供鼠干细胞，实验组受鼠回输供鼠干细胞＋脐血单个核细胞。于移植后＋7 d、＋14 d、＋21 d、＋28 d和＋50 d对存活受鼠进行断尾采血，检测白细胞、血红蛋白和血小板，分析造血重建情况。处死小鼠时取外周血1 mL，采用荧光原位杂交技术检测Y染色体比例，计算嵌合率。两组受鼠各时间点白细胞、血红蛋白及血小板检测结果采用成组t检验进行比较，嵌合率采用卡方检验进行比较。P<0.05为差异有统计学意义。

结果

移植后＋14 d实验组和对照组白细胞数量分别为(8.4±2.6)×109/L和(3.8±1.0)×109/L，差异有统计学意义(t＝6.968，P<0.05)；其他时间点两组白细胞数量差异无统计学意义。移植后两组受鼠各时间点血红蛋白检测结果差异均无统计学意义。移植后＋7 d实验组和对照组血小板数量分别为(125±40)×109/L和(64±15)×109/L，移植后＋14 d分别为(282±47)×109/L和(163±41)×109/L，差异均有统计学意义(t＝6.366和8.093，P均<0.05)；其他时间点两组血小板数量差异无统计学意义。

结论

使用脐血细胞作为第三方细胞辅助输注后，可促进单倍型造血干细胞移植受鼠早期白细胞和血小板较快恢复。

关键词: 单倍型, 造血干细胞移植, 脐血, 造血重建, 小鼠
Objective

To analyze the hematopoietic reconstitution in the mouse model of haploidentical hematopoietic stem cell transplantation when using umbilical cord blood cells as the third party cells.

Methods

The CB6/F1 mice were selected as the donor and the BALB/C mice were selected as the receptor. All the recipient mice were infused with donor stem cells after myeloablative with Co60 to establish the mouse model of the haploidentical hematopoietic stem cell transplantation. Forty recipient mice were equally divided into the experimental group and the control group. The mice in the control group were infused with donor mice-derrived haploidentical hematopoietic stem cell, and the mice in the experimental group were infused with umbilical cord blood cells and donor mice-derrived haploidentical hematopoietic stem cell. Blood collection by tail snipping in the viable recipient mice was performed at intervals of + 7, + 14, + 21, + 28, + 50 days after transplantation and then the assay of white blood cell, hemoglobin and blood platelet were performed to evaluate the hematopoietic reconstitution. One mL peripheral blood was obtained when the mice were sacrificed. The proportion of Y chromosome was dected by fluorescence in situ hybridization and then the proportion of chimerism was calculated. The test results of white blood cell, hemoglobin and blood platelet between the 2 groups were compared by t test, and the proportion of chimerism were compared by chi-square test. P<0.05 was considered statistically significant.

Results

The white blood cell counts at + 14 days after transplantation between the experimental group and the control group were (8.4±2.6)×109/L and (3.8±1.0)×109/L respectively, which had statistical significance (t=6.968, P<0.05), while no statistical signicance was found at other time points. No statistical significance was found between the 2 groups for hemoglobin. The blood platelet counts at + 7d, + 14d after transplantation between the experimental group and the control group were (125±40)×109/L and (64±15)×109/L, (282±47)×109/L and (163±41)×109/L respectively, which had statistical significance (t=6.366 and 8.093, P<0.05), while no statistical signicance was found at other time points.

Conclusions

The recovery of peripheral blood white blood cell and blood platelet in the mouse model of haploidentical hematopoietic stem cell transplantation was faster with the help of umbilical cord blood cells infusion.

Key words: Haplotype, Hematopoietic stem cell transplantation, Umolbilical cord blood, Hematopoietic reconstitution, Mouse
表1 单倍型造血干细胞移植＋7 d两组受鼠白细胞、血红蛋白和血小板检测结果(±s)
组别 例数 白细胞(×109/L) 血红蛋白(g/L) 血小板(×109/L)
实验组 20 1.1±0.3 186±14 125±40
对照组 20 1.0±0.3 189±14 64±15
t值 － 1.246 －0.728 6.366
P值 － >0.05 >0.05 <0.05
表2 单倍型造血干细胞移植＋14 d两组受鼠白细胞、血红蛋白和血小板检测结果(±s)
组别 例数 白细胞(×109/L) 血红蛋白(g/L) 血小板(×109/L)
实验组 18 8.4±2.6 176±21 282±47
对照组 18 3.8±1.0 182±19 163±41
t值 － 6.968 －0.918 8.093
P值 － <0.05 >0.05 <0.05
表3 单倍型造血干细胞移植＋21 d两组受鼠白细胞、血红蛋白和血小板检测结果(±s)
组别 例数 白细胞(×109/L) 血红蛋白(g/L) 血小板(×109/L)
实验组 16 8.9±6.5 174±24 899±243
对照组 15 8.3±1.9 180±16 939±259
t值 － 0.369 －0.842 －0.449
P值 － >0.05 >0.05 >0.05
表4 单倍型造血干细胞移植＋28 d两组受鼠白细胞、血红蛋白和血小板检测结果(±s)
组别 例数 白细胞(×109/L) 血红蛋白(g/L) 血小板(×109/L)
实验组 14 10.5±2.1 159±33 846±500
对照组 11 8.4±3.8 156±26 631±369
t值 － 1.789 0.150 1.192
P值 － >0.05 >0.05 >0.05
表5 单倍型造血干细胞移植＋50 d两组受鼠白细胞、血红蛋白和血小板检测结果(±s)
组别 例数 白细胞(×109/L) 血红蛋白(g/L) 血小板(×109/L)
实验组 11 13.1±5.5 164±6 512±348
对照组 7 12.6±3.4 168±8 605±410
t值 － 0.234 －0.320 －0.519
P值 － >0.05 >0.05 >0.05
1
Czerw T, Labopin M, Schmid C, et al. High CD3＋ and CD34＋ peripheral blood stem cell grafts content is associated with increased risk of graft-versus-host disease without beneficial effect on disease control after reduced-intensity conditioning allogeneic transplantation from matched unrelated donors for acute myeloid leukemia - an analysis from the Acute Leukemia Working Party of the European Society for Blood and Marrow Transplantation[J]. Oncotarget, 2016，7(19):27255-27266.
2
杜振兰，陈鹏，罗荣牡，等. 第三方脐血辅助输注的单倍型造血干细胞移植36例疗效分析[J/CD]. 中华临床医师杂志：电子版，2014，8(6):1200-1203.
3
蔡芳芳，俞康，江松福. 小鼠同种异基因骨髓移植aGVHD动物模型的建立[J]. 温州医学院学报，2009，39(4):320-324.
4
扈江伟，金建刚，王莉莎，等. 异基因骨髓移植小鼠GVHD模型的建立[J]. 军事医学科学院院刊，2007，31(2):141-144.
5
汤永永，唐颖，张玉华，等. 基于CFSE染色的小鼠淋巴细胞体内示踪方法的建立[J]．中国输血杂志，2011，24(10):850-853.
6
Las′ovicka J, Budinský V, Spísek R, et al. Assessment of lymphocyte proliferation: CFSE kills dividing cells and modulates expression of activation markers[J]. Cellular Immunol, 2009, 256(1-2):79-85.
7
Lee SJ. Have we made progress in the management of chronic graft-vs-host disease？[J]. Best Pract Res Clin Haematol, 2010，23(4):529-535.
8
Yang J, Fan H, Hao J, et al. Amelioration of acute graft-versus-host disease by adoptive transfer of ex vivo expanded human cord blood CD4＋CD25＋ forkhead box protein 3＋ regulatory T cells is associated with the polarization of Treg/Th17 balance in a mouse model[J]. Transfusion, 2012，52(6):1333-1347.
9
Patel A, Gruber P. Severe infections in neutropenic patients[J]. Curr Opin Crit Care, 2015，21(6):586-592.
10
Abu-Khader A, Law KW, Jahan S, et al. Paracrine factors released by osteoblasts provide strong platelet engraftment properties[J]. Stem Cells, 2019，37(3):345-356.
11
Goto T, Murata M. Mesenchymal stem cell therapy in hematopoietic stem cell transplantation[J]. Rinsho Ketsueki, 2018，59(2):195-204.
12
Vavilov VN, Averyanova My, Bondarenko SN, et al. Bacterial infections in the early period after allogeneic bone marrow transplantation[J]. Ter Arkh, 2015，87(7):88-93.
13
Wu QL, Liu XY, Nie DM, et al. Umbilical cord blood-derived mesenchymal stem cells ameliorate graft-versus-host disease following allogeneic hematopoietic stem cell transplantation through multiple immunoregulations[J]. J Huazhong Univ Sci Technolog Med Sci, 2015，35(4):477-484.
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