神经母细胞瘤细胞Boyden小室体外侵袭模型的建立及意义探讨

Establishing Boyden chamber for invasive neuroblastoma

目的 建立神经母细胞瘤细胞Boyden小室体外侵袭模型.对其实验条件、方法 及模型的意义进行探讨.方法 将不同浓度（1.5×105、2.0×105、2.5×105、3.0×105/ml）的100μl 4组神经母细胞瘤细胞系置入Boyden小室侵袭模型上室,分别培养24、36、48、60h,观察不同浓度的细胞在不同时间穿透基质胶聚碳酸酯膜的细胞数,用Boyden小室测定4组神经母细胞瘤细胞系侵袭的能力.结果 Boyden小室侵袭模型显示4组不同细胞系的侵袭能力不完全相同,在第24 h透过ECM胶聚碳酸酯膜的细胞较少,随时间的延长而增多,在48h前后增幅最明显;同时,随细胞浓度的增高,透过细胞数也增加,但细胞浓度大于2.5×105/ml时,透过ECM胶聚碳酸酯膜的细胞数变化不显著;在同一条件下,有骨髓转移能力的细胞系其侵袭能力大于无骨髓转移能力的细胞系.结论 本实验成功建立了神经母细胞瘤细胞Boyden小室体外侵袭模型,该模型对体外神经母细胞瘤细胞侵袭能力的测定和研究及对探明神经母细胞瘤的转移机制和影响因素具有重要意义.

Objective To establish a Boyden chamber for invasive human neuroblastoma cell study. Methods We used Boyden chamber in vitro assay to investigate the invasion of four cell lines.100μl four cell suspension of various concentrations （1.5 × 105、 2. 0 × 105 、 2. 5 × 105 、 3. 0 × 105/ml）was added into the upper chamber of the Boyden chamber,and the cells were allowed to penetrate the membrane for 24,36,48 and 60 hours respectively. The invasion rate was assessed by quantifying cells on the under surface of the membrane. Results Boyden chamber invasion models showed that the invasion of four different cell lines was different. The invasive cell quantity was low 24 hours after the incubation, and it increased dramatically at 48 hours. After 48 hours the rate reached a relative plateau. The invasion quantity was high as the cells concentration added increased. The increase slowed down when the cell concentration was above 2. 5 × 105/ml. Under the same conditions, the invasion ability of cells with the capacity of bone marrow metastasis than the cells without ability of bone marrow metastasis.Conclusions We successfully adopted the Boyden chamber for the neuroblastoma cells. It will be crucial for future study of the invasive ability of the neuroblastoma cells.