中国组织工程研究

中国组织工程研究 ›› 2010, Vol. 14 ›› Issue (19): 3550-3553.doi: 10.3969/j.issn.1673-8225.2010.19.026

• 干细胞基础实验 basic experiments of stem cells • 上一篇    下一篇

应用含标记基因K562细胞制备的小鼠白血病模型

王存邦，白 海，葸 瑞，钱 震，张 茜   

  • 出版日期:2010-05-07 发布日期:2010-05-07
  • 作者简介:Wang Cun-bang, Doctor, Associate chief physician, Master’s supervisor, Hematology Center, General Hospital of Lanzhou Military Area Command of Chinese PLA, Lanzhou 730050, Gansu Province, China wangcunbang98@163.com
  • 基金资助:

    全军“十一五”杰出人才基金资助项目（06J005），
    兰州军区医药科研基金（LXH-2007006）资助。

Establishment of leukemia mouse models using marker gene-containing K562cell line

Wang Cun-bang, Bai Hai, Xi Rui, Qian Zhen, Zhang Qian   

  1. Hematology Center, General Hospital of Lanzhou Military Area Command of Chinese PLA, Lanzhou 730050, Gansu Province, China
  • Online:2010-05-07 Published:2010-05-07
  • About author:王存邦，男，1966年生，青海省西宁市人，汉族，1997年解放军第四军医大学毕业，博士，副主任医师，硕士生导师，主要从事造血干细胞移植研究。 Wangcunbang98＠163.com
  • Supported by:

    the Outstanding Talent Foundation of the Whole Army during the 11th Five-Year Plan Period, No. 06J005*;
    the Medicine Scientific Research Foundation of Lanzhou Military Area Command of Chinese PLA, No. LXH-2007006*

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被引次数

8

摘要：

背景：文献报道应用NOD/SCID及SCID小鼠可以建立移植性人白血病小鼠模型，但由于小鼠存在免疫缺陷，使得在自体干细胞移植及移植后相关过继免疫治疗方面的研究受到限制和影响。

目的：探索用SPF级Balb/c小鼠和转染GFP及NeoR基因的K562细胞株制备白血病模型的方法。

方法：实验分为5组，A、B组和C、D组分别经X射线照射2 Gy和3 Gy，24 h后取对数生长期的K562(GFP+/Neo+) 细胞， A、C组尾静脉注射2×106个/只；B、D组尾静脉注射5×106个/只；E组为正常对照组。观察小鼠生存时间，进行骨髓细胞及外周血白细胞分类，采用流式细胞仪测定GFP阳性细胞及PCR方法测定Neo基因。

结果与结论：实验各组在接种5~7 d时发病，分别于30，23，24，17 d内全部死亡；生存天均显著短于正常对照组(P < 0.01)。  体质量均较正常对照组显著下降(P < 0.05)。小鼠的白血病发病率为100%，无自发缓解。随接种细胞数量的增加和照射剂量的增加，小鼠的存活时间缩短，且外周血及骨髓中白血病细胞所占比例增加。流式细胞仪测定及PCR方法也证实了GFP+细胞和NeoR基因在肝、脾中的存在。结果证实 Balb/c小鼠经照射后从尾静脉注射K562细胞株可以制备获得白血病小鼠模型。

关键词: 绿色荧光蛋白基因, K562细胞, 新霉素磷酸转移酶基因, 白血病小鼠模型, Balb/c小鼠

Abstract:

BACKGROUND: Leukemia mouse models were established using NOD/SCID and SCID mice. However, mice have immune deficiencies, which limit and affect the research of adoptive immunotherapy during and following autologous hematopoietic stem cell transplantation.

OBJECTIVE: To explore the method of preparing leukemia mouse models using SPF grade Balb/c mouse and K562 cell line transfected with green fluorescent protein (GFP) and NeoR genes.

METHODS: This study established five groups. Groups A and B as well as groups C and D were irradiated 2Gy or 3Gy for 24 hours respectively, and then injected with K562(GFP+/Neo+) cells in exponentiall growth period at a density of 2×106 /mouse or 5×106 /mouse via vena caudalis. Group E served as normal control group. Survival time of mice was observed. Bone marrow cells and peripheral blood leucocytes were classified. GFP-positive cells and Neo gene were respectively determined using flow cytometry and PCR.

RESULTS AND CONCLUSION: Mice of experimental group fell ill 5-7 days later. All of mice died within 30, 23, 24, 17 days respectively. Survival days were significantly shorter compared with the normal control group (P < 0.01). The weight decreased significantly than that in normal control group (P < 0.05). The incidence of leukemia in processed mice was 100%, and had no natural relief. With the increase of infused cell population and exposure dose, survival days became short, and cell proportion in peripheral blood and bone marrow increased gradually. Flow cytometry and PCR have confirmed the existence of GFP and NeoR gene in the live and spleen. Results verified that the leukemia mouse model can be made by infusing K562 cells by vena caudalis into Balb/c mice after irradiation.

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