摘要
目的:应用无血清培养液(SFM)培养Lovo细胞系,克隆分离具有干细胞样特性SP细胞.方法:应用SFM培养Lovo细胞,分离成球样生长的细胞群作为SP细胞,并通过有限稀释,分化,自我更新,含血清培养基(SSM)和SFM交替培养及Musashi-1化学染色方法来鉴定SP细胞.结果:Lovo细胞中约含有0.54%-0.62%的SP能够在无血清培养基中能够存活、增殖,形成悬浮的肿瘤细胞球;在SFM中加入血清可促使SP细胞分化;SP细胞可以连续传代,并可交替培养于SSM和SFM中,细胞形态无明显变化;SP细胞Musashi-1染色阳性.结论:应用SFM可从Lovo细胞中分离出极少量的具有干细胞特性的SP细胞.
AIM： To investigate whether coloncancer cell line Lovo contains side population （SP） with cancer stern 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.
CONCLUSION： Lovo cell line contains a tiny minority of SP cells with stem cell properties that can be maintained in SFM using a floatingculture method for a long time.
出处
《世界华人消化杂志》
CAS
北大核心
2007年第9期953-959,共7页
World Chinese Journal of Digestology
关键词
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
Lovo colon cancer cell line
Side population
Tumor stem cells
Serum-free culture medium
Floating-culture system
