Huang Qiang, Dong Jun, Zhu Yu-de, Zhang Quan-bin, Ji Xiao-yan, Wang Ai-dong, Lan Qing
Neurosurgery Department and Brain Tumor Research Laboratory, 2nd Affiliated Hospital of Suzhou University, Suzhou 215004, China.
Zhonghua Zhong Liu Za Zhi. 2006 May;28(5):331-3.
To isolate and culture tumor stem cells from glioma tissues obtained at surgical operation and to study their biological characteristics.
Glioma tissues obtained from surgically resected specimens of 8 patients were fully chopped, trypsinized, and filtered to prepare single cell suspensions. The cells were cultured in serum-free medium with EGF, LIF and bFGF. CD133(+) cells were purified by magnetic cell sorting, and cultured continuously in vitro to obtain tumor cell spheres. Tumor stem cells of the 5th passage were induced to differentiate with 10% FBS, and expression of cell differentiation markers such as Nestin, MAP2, GFAP was evaluated with immunocytochemistry techniques.
CD133(+) cells were successfully separated and cultured from one anasplastic mixed astrocyte-ependymocyte type glioma specimen. These cells maintained a sphere-like growth status in vitro (3 months, 14 passages), and can self-renew, proliferate and conditionally differentiate into MAP2(+) and GFAP(+) cells. However, CD133(-) cells did not possess these properties.
Glioma tissue contains tumor stem cells. Those cells can be cultured and passaged in vitro for a long term, and therefore to offer new approaches for studying cellular and molecular biology of glioma.
从手术切除的胶质瘤组织中分离培养肿瘤干细胞，并研究其生物学特性。
将8例手术切除标本中获取的胶质瘤组织充分切碎，经胰蛋白酶消化、过滤制成单细胞悬液。细胞在含表皮生长因子（EGF）、白血病抑制因子（LIF）和碱性成纤维细胞生长因子（bFGF）的无血清培养基中培养。通过磁珠细胞分选法纯化CD133(+)细胞，并在体外持续培养以获得肿瘤细胞球。第5代肿瘤干细胞用10%胎牛血清诱导分化，采用免疫细胞化学技术评估神经巢蛋白（Nestin）、微管相关蛋白2（MAP2）、胶质纤维酸性蛋白（GFAP）等细胞分化标志物的表达。
成功从1例间变性混合性星形-室管膜细胞型胶质瘤标本中分离培养出CD133(+)细胞。这些细胞在体外保持球形生长状态（3个月，传代14次），具有自我更新、增殖能力，并可诱导分化为MAP2(+)和GFAP(+)细胞。而CD133(-)细胞不具备这些特性。
胶质瘤组织中含有肿瘤干细胞。这些细胞能够在体外长期培养传代，为胶质瘤细胞和分子生物学研究提供了新途径。
