Chinese Journal of Tissue Engineering Research ›› 2010, Vol. 14 ›› Issue (19): 3550-3553.doi: 10.3969/j.issn.1673-8225.2010.19.026

Previous Articles     Next Articles

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*

PDF

432

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.

CLC Number: