斑马鱼异种异体急性髓系白血病模型的建立

Establishment of a Xenografted Acute Myeloid Leukemia Model by using Zebrafish

目的:探讨利用人急性髓系白血病干细胞和斑马鱼建立斑马鱼白血病模型,为白血病的研究及治疗药物的筛选提供更为直接的体外研究模型和体内实验依据。方法:利用MitoRed染料对白血病干样细胞KG-1a细胞进行标记,然后通过显微注射法把KG-1a细胞植入斑马鱼胚胎的卵黄囊中,观察移植后斑马鱼胚胎的大体存活状态、细胞在斑马鱼体内的增殖和扩散转移;应用组织学切片观察移植后细胞对斑马鱼组织器官的侵袭作用。结果:KG-1a细胞植入斑马鱼胚胎的卵黄囊后,MitoRed标记的KG-1a细胞数目随时间延长而增多并逐渐扩散至整个腹腔,体外计数结果也显示植入细胞明显增殖,表明植入的白血病干细胞能够在斑马鱼体内存活,增殖并扩散。进一步的研究显示,在斑马鱼肝脏组织发现有白血病细胞的浸润,HE染色提示浸润细胞具有KG-1a细胞核型特征。结论:异种异体的人急性髓系白血病干细胞KG-1a可以植入斑马鱼体内并成功建立斑马鱼的白血病模型。该模型的建立可为白血病的研究及治疗药物的筛选提供一种更为直接高效的体外研究平台。

Objectve：To investigate the feasibility of establishing xenografted leukemia model by zebrafish,so as to provide the more direct model in vitro and experimental evidence for study of acute myeloid leukemia and screening of the drugs for targeting therapy.Methods：Acute myeloid leukemia cell line KG-1α was labeled with red fluorescent dyeMitoRed,then the labeled cells were injected into the yolk sac of zebrafish embryos.Morphological observation,cell count and histopathological detection were used to analyse the infiltration and metastasis of KG-1a cells in zebrafish.Results：KG1a cells could proliferate and gradually spread to the entire abdominal cavity of the zebrafish after KG-la cells were injected into the yolk sac during 1- 7,the results of cell counting in vitro also proved a significant proliferation of KG-1a cells in zebrafish,suggesting that the implanted leukemia stem cells could survive,proliferate and spread in zebrafish.Further study showed that the implanted cells could be transfered to the liver of zebrafish,these cells displayed the signature of KG-la cells by hematoxylin- eosin（HE） staining.Conclusions：Human acute myeloid leukemia cells KGla can survive,proliferate and migrate in zebrafish,suggesting xenografted leukemia model of zebrafish has been successfully established.This model may be benefitcial for the study of acute myeloid leukemia and the screening of the drugs for targeting merapy of acute myeloid leukemia.