上海交通大学学报（医学版）

上海交通大学学报（医学版） ›› 2019, Vol. 39 ›› Issue (10): 1110-.doi: 10.3969/j.issn.1674-8115.2019.10.002

• 论著·基础研究 • 上一篇    下一篇

小鼠胎肝单个核细胞行骨髓腔内注射重建造血系统的效果

伊庆福1，迟亚男1，刘光辉2, 3，杨冠恒2, 3，龚秀丽2, 3，范书玥1，曾凡一1, 2, 3   

  1. 1.上海交通大学基础医学院组织胚胎学与遗传发育学系，上海 200025；2.上海交通大学附属儿童医院，上海交通大学医学遗传研究所，上海 200040；3. 卫生部医学胚胎分子生物学重点实验室暨上海市胚胎与生殖工程重点实验室，上海 200040
  • 出版日期:2019-10-28 发布日期:2019-11-22
  • 作者简介:伊庆福（1989—），男，硕士生；电子信箱：18101925435@163.com。
  • 基金资助:
    国家重大科学研究计划（2014CB964700）；上海市自然科学基金（16ZR1428600）

Intra-bone marrow injection of fetal liver mononuclear cells to reconstruct the hematopoietic system in mice

YI Qing-fu1, CHI Ya-nan1, LIU Guang-hui2, 3, YANG Guan-heng2, 3, GONG Xiu-li2, 3, FAN Shu-yue1, ZENG Fan-yi1, 2, 3   

  1. 1. Department of Histoembryology, Genetics & Development, Shanghai Jiao Tong University College of Basic Medical Sciences, Shanghai 200025, China; 2. Shanghai Institute of Medical Genetics, Shanghai Childrens Hospital, Shanghai Jiao Tong University, Shanghai 200040, China; 3. Key Laboratory of Embryo Molecular Biology, Ministry of Health; Shanghai Key Laboratory of Embryo and Reproduction Engineering, Shanghai 200040, China
  • Online:2019-10-28 Published:2019-11-22
  • Supported by:
    National Major Scientific Research Plan, 2014CB964700; Natural Science Foundation of Shanghai, 16ZR1428600

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摘要： 目的·观察用小鼠胎肝单个核细胞悬液行骨髓腔内注射以重建小鼠造血系统的效果。方法·用Ficoll分离技术获取胎龄13.5 d的小鼠胎肝单个核细胞悬液，采用磁珠分选的方法获得造血干细胞（hematopoietic stem cell，HSC）并计数。以骨髓腔内注射的方法对137Cs辐射清髓后的小鼠胫骨进行小鼠胎肝单个核细胞移植。对移植后的受体小鼠通过血常规检测、血涂片及骨髓涂片镜检的方法观察造血系统的重建情况。对受体小鼠移植后1年骨髓中的供体HSC进行流式细胞检测。结果·胎龄13.5 d的小鼠胎肝单个核细胞中HSC占0.171%。小鼠胎肝单个核细胞移植后1周，外周血即能探测到供体细胞来源的血细胞；移植后第3周受体小鼠外周血白细胞、红细胞、血红蛋白基本恢复至辐照前水平；第5周外周血中供体细胞来源的血细胞比例趋于稳定；移植后1年受体小鼠骨髓中的HSC基本来自供体细胞。结论·小鼠胎肝单个核细胞通过骨髓腔内注射的移植方法能高效重建小鼠造血系统。

关键词: 造血干细胞, 骨髓腔内注射, 胎肝, 小鼠

Abstract:

Objective · To observe the effect of intra-bone marrow injection (IBMI) of fetal liver mononuclear cell suspension on reconstructing the hematopoietic system in mice. Methods · The fetal liver mononuclear cells the mice with gestational age of 13.5 days were obtainedFicoll separation technique, and then the hematopoietic stem cells (HSCs) were obtainedmagnetic bead sorting. The fetal liver mononuclear cells were injected into the tibias of 137Cs irradiated miceIBMI. The recipient mice were observed for reconstitution of the hematopoietic systemroutine blood examination, blood smear and bone marrow smear. The donor HSCs in the bone marrow of the recipient mice was subjected to flow cytometry 1 year after transplantation. Results · HSCs accounted for 0.171% in the mofetal liver mononuclear cells. One week after transplantation of mofetal liver mononuclear cells, blood cells derived donor cells were detected peripheral blood; in the third week after transplantation, peripheral blood leukocytes, red blood cells and hemoglobin of recipient mice were restored to the pre-irradiation levels; the percentage of donor-derived blood cells in peripheral blood in the fifth week was stable; the HSCs in the bone marrow of recipient mice were derived donor cells 1 year after transplantation. Conclusion · The murine fetal liver mononuclear cells can efficiently reconstruct the hematopoietic system of miceIBMI.

Key words: hematopoietic stem cell (HSC), intra-bone marrow injection (IBMI), fetal liver, mouse

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