大肠癌细胞系Lovo中亚群(SP)细胞的分离培养和鉴定

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.

Isolation and identification of side population in Lovo colon cancer cell line

AIM:To investigate whether coloncancer 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,alterna- tive 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 pro- liferative,self-renewal and differentiation po- tential,which were responsible for the floating tumor clone.Serum addition into SFM resulted in the proliferation of SP cells;after several gen- erations 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 floating- culture method for a long time.