目的探讨有效的实体性造釉细胞型颅咽管瘤细胞原代培养方法,检测体外培养的颅咽管瘤细胞生物学特性。方法采用胰酶消化法建立实体性颅咽管瘤细胞株,并通过滤膜过滤纯化细胞;倒置显微镜、HE染色观察生长情况及细胞形态,免疫组化证实其细胞来源,噻唑蓝比色(MTT)法绘制细胞生长曲线,并用流式细胞仪分析细胞周期。结果肿瘤呈典型上皮样形态,免疫细胞鉴定颅咽管瘤细胞原代培养成功,肿瘤细胞增殖速度缓慢,且均为二倍体细胞。结论造釉细胞型颅咽管瘤细胞通过酶消化及改良的血清培养基法成功建立,且滤膜过滤可以提高所培养细胞的纯度和成功率。
Objective To establish an efficient method of primary solid adamantinomatous craniopharyngioma cells culture and observe the behaviour of ceils in vitro. Methods The specimen of solid adamantinomatous craniopharyngioma were treated with trypsin and cultured in vitro, and purified by filter membrane. The cell morphology was observed by inverted microscope and hematoxylin eosin staining. The expression of Pan - ek was examined by immunocytochemistry. The MTT assay and flow cytometry were employed to study the cell growth and proliferation. Results The cell morphology was similar to that of typical epithelial cell and the staining of Pan - ck was positive in immunocytochemistry. The cultured tumor cells grew slowly and were all diploid. Conclulions The adamantinomatous craniopharyngioma cells could be obtained by enzymatic digestion method and special culture media. Filter membrane could be applied to improve the purity and success rate of craniopharyngioma cells.