大肠癌细胞系Lovo中亚群(SP)细胞的分离培养和鉴定
Lovo大肠癌细胞系;细胞亚群;肿瘤干细胞;悬浮培养;无血清培养基,王锡山,刘彦龙,杨艳梅,王锡山,通讯作者:,IsolationandidentificationofsidepopulationinLovocoloncancerc
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王锡山, 刘彦龙, 哈尔滨医科大学附属肿瘤医院腹外科 黑龙江省哈尔滨市 150040
杨艳梅, 黑龙江省肿瘤研究所 黑龙江省哈尔滨市 150040
王锡山, 教授, 博士生导师, 主要从事腹部肿瘤的基础研究和外科治疗.
通讯作者: 王锡山, 150040, 哈尔滨市南岗区哈平路150号, 哈尔滨医科大学附属肿瘤医院腹外科. wxshan1208@yahoo.com.cn
电话: 0451-86298898
收稿日期: 2007-01-23 接受日期: 2007-01-31
Isolation and identification of side population in Lovo colon cancer cell line
Xi-Shan Wang, Yan-Long Liu, Yan-Mei Yang
Xi-Shan Wang, Yan-Long Liu,Department of Abdominal Surgery, the Affliliated Tumor Hospital of Harbin Medical University, Harbin 150040, Heilongjiang Province, China
Yan-Mei Yang,Institute of Tumor research, Harbin 150040, Heilongjiang Province, China
Correspondence to:Xi-Shan Wang, Department of Abdominal Surgery, the Affiliated Tumor Hospital of Harbin Medical University, 150 Haping Road, Nangang District, Harbin 150040, Heilongjiang Province, China. wxshan1208@yahoo.com.cn
Received:2007-01-23 Accepted:2007-01-31
Abstract
AIM: To investigate whether colon cancer cell line Lovo contains side population (SP) with cancer stem cell-like properties.
METHODS: Lovo cell line was cultivated in serum-free medium and SP cells reforming into floating spheres were isolated. The isolated SP cells were identified by limiting-dilution assay, differentiation assay, self-renewal assay, alternative cultivation assay and immunocytochemical staining with anti-Mus-1 assay.
RESULTS: In the absence of serum, a minority (0.54%-0.62%) of SP cells in Lovo cells survived, proliferated and assembled into the suspended tumor cell spheres. Lovo SP cells possessed proliferative, self-renewal and differentiation potential, which were responsible for the floating tumor clone. Serum addition into SFM resulted in the proliferation of SP cells; after several generations and alternated cultivation in SSM and SFM, SP cells maintained their characters. The stem cell surface marker Mus-1 was observed in Lovo SP cells after Musashi-1 staining ......
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