摘要
目的:从人恶性胶质瘤中分离并鉴定胶质瘤干细胞。方法:采用神经干细胞无血清培养方法,分离胶质瘤干细胞,免疫细胞化学法、有限梯度稀释实验、二代球体形成分析、流式细胞分析等方法对胶质瘤干细胞进行鉴定。结果:3例患者原代培养胶质瘤细胞中CD133+细胞分别为7.4%、2.8%和4.2%。每100个原代胶质瘤细胞中可形成1个左右的胶质瘤细胞球;胶质瘤细胞球细胞均表达神经干细胞标记CD133和Nestin,不表达分化标志GFAP和MAP2,少数细胞表达分化标记MBP。每100个原代胶质瘤细胞球体细胞可形成6.07±2.15个悬浮生长的二代胶质瘤细胞球,二代细胞球细胞表达CD133和Nestin。胶质瘤细胞球细胞分化后细胞多数表达GFAP,少数表达MAP2和MBP。胶质瘤球体干细胞的增殖较原代胶质瘤细胞慢。胶质瘤球体细胞1×103个时可成瘤,原代培养的胶质瘤细胞需1×107。结论:采用神经干细胞培养条件可以很简便的分离出悬浮生长的胶质瘤球体干细胞。分离出的胶质瘤球体干细胞表达CD133抗原,具备肿瘤干细胞的基本特征。
OBJECTIVE： To isolate and identificate the brain tumor stem cells （BTSCs） within human gliomas tissue in vitro. METHODS： By switching primary human glioblastoma cells into a defined serum-free neural stem cells medium, glioblastoma spheres were isolated. By limiting dilution assay, secondary spheres forming assay, flow cytometry assay and so on, BTSCs were identificated. RESULTS： Glioblastoma spheres consisted of BTSCs expressing CD133 and nestin but no GFAP and MAP2, and a few BTSCs expressed MBP. The percentage of CD133^＋ cell in 3 cases of primary cultured glioma cells were 7.4%, 2.8% and 4.2 %. About 1% cells in primary cultured glioma cells generated free-floating glioblastoma spheres and 100 cells of primary tumor spheres generated 6.07±2. 15 secondary glioma spheres expressing CD133 and Nestin. Most cells produced from BTSC spheres expressed GFAP, and a few expressed MAP2 and MBP. Glioma BTSCs cultured in serum-free neural stem cells medium showed a lower degree of cell proliferation than primary glioma cells cultured in medium with 10% FBS. BTSCs had stronger capable of tumor initiation in nude mice than primary cultured glioma cells. CONCLUSIONS： CD133^＋ BTSCs can be isolated from primary cultured glioma cells by cultured in serum-free neural stem cells medium. BTSCs express CD133 antigen and have characteristics of cancer stem cells.
出处
《中华肿瘤防治杂志》
CAS
2007年第13期972-975,共4页
Chinese Journal of Cancer Prevention and Treatment
