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目的探讨胰腺癌细胞在二维平面培养和三维培养(Ⅰ型胶原和细胞外基质胶)中的生长特性。方法将3株胰腺癌细胞株SW1990、PCT和ASPC-1分别采用上述3种培养方式进行培养,观察细胞生长形态。采用CCK-8法测定细胞生长曲线,采用乙醇固定碘化丙锭染色法检测细胞周期分布。结果细胞在二维平面培养系统中呈单层贴壁生长;在Ⅰ型胶原和细胞外基质胶中细胞形成多细胞球样体(multicellular spheroid,MCS),其生长速度较二维平面培养中的细胞慢。在二维平面培养中生长的SW1990、PCT和ASPC-1细胞的S期细胞的比例分别为(29.6±3.0)%、(33.6±2.1)%和(33.1±1.8)%,明显高于在Ⅰ型胶原中培养4 d和8 d形成的MCS的S期细胞比例〔(18.2±5.1)%、(14.5±3.2)%和(24.7±2.6)%〕,P<0.05,而G2/M期的细胞比例差异没有统计学意义(P>0.05)。在Ⅰ型胶原中培养4 d和8 d的SW1990和PCT细胞以及培养8 d的ASPC-1细胞的G0/G1期细胞比例明显高于其在二维平面培养中的细胞比例(P<0.05)。在Ⅰ型胶原中培养4 d的ASPC-1细胞和SW1990细胞的S期细胞比例明显高于其培养8 d的细胞比例(P<0.05)。结论不同的培养方式、培养介质对胰腺癌细胞的生长具有较大的影响。MCS三维培养系统能更好地模拟体内肿瘤细胞的生长状况。
Objective To investigate the growth characteristics of pancreatic cancer cells in two-dimensional planar culture and three-dimensional culture (type I collagen and extracellular matrix glue). Methods The three pancreatic cancer cell lines SW1990, PCT and ASPC-1 were cultured by the above three culture methods, and the cell growth morphology was observed. The cell growth curve was determined by CCK-8 method, and the cell cycle distribution was detected by propidium iodide staining. Results The cells grew monolayer in two-dimensional planar culture system. In type I collagen and extracellular matrix gel, the cells formed multicellular spheroid (MCS) which grew faster than those in 2D planar culture Slow cells. The proportion of S phase cells in SW1990, PCT and ASPC-1 cells grown in two-dimensional planar culture were (29.6 ± 3.0)%, (33.6 ± 2.1)% and (33.1 ± 1.8)%, respectively, (18.2 ± 5.1)%, (14.5 ± 3.2)% and (24.7 ± 2.6)%, respectively) of MCS formed on day 4 and day 8, P <0.05, while the percentage of cells in G2 / M phase No significant difference in the proportion of cells (P> 0.05). The percentage of G0 / G1 phase cells in SW1990 and PCT cells cultured for 4 d and 8 d on type Ⅰ collagen and ASPC-1 cells cultured for 8 d was significantly higher than that in 2D planar culture (P <0.05) . The proportion of S phase cells in ASPC-1 cells and SW1990 cells cultured for 4 d in type Ⅰ collagen was significantly higher than that in cultured cells for 8 d (P <0.05). Conclusion Different culture methods and media have a greater impact on the growth of pancreatic cancer cells. MCS three-dimensional culture system can better simulate the growth of tumor cells in vivo.