胰腺癌细胞在二维和三维培养系统中的生长特性研究

Growth Characteristics of Pancreatic Cancer Cells in Two-or Three-Dimensional Culture Systems

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 the two-dimensional culture system（monolayer） and three-dimensional culture system（type Ⅰ collagen and extracellular matrix gel）.Methods Three pancreatic cancer cell lines（SW1990,PCT,and ASPC-1） were cultured in monolayer,type Ⅰ collagen,and extracellular matrix gel,respectively.The growth patterns were observed,growth curves were detected by CCK-8 test,and the cell cycle distributions were analyzed by propidium iodide staining.Results In the two-dimensional culture system,cells grew in monolayer.In the type Ⅰ collagen and the ECM gel three-dimensional culture system,cells formed multicellular spheroids（MCS）,of which the growth rates were slower than those of the cells in monolayer.The proportions of S phase of SW1990,PCT,and ASPC-1 cells in two-dimensional culture system were significantly more than those in the type Ⅰ collagen on 4 d and 8 d（（29.6±3.0）% vs.（18.2±5.1）%,（33.6±2.1）% vs.（14.5±3.2）%,（33.1±1.8）% vs.（24.7±2.6）%;P〈0.05）,while the difference of proportion of three cell lines in G2/M phase was not different between two-dimensional culture system and type Ⅰ collagen（P〈0.05）.The proportions of G0/G1 phase of SW1990 and PCT cells cultured in the type Ⅰ collagen on 4 d and 8 d and ASPC-1 cells cultured in the type Ⅰ collagen on 4 d were significant more than those cultured in two-dimensional culture system（P〈0.05）.The proportions of S phase of ASPC-1 cells and SW1990 cells cultured in the type Ⅰ collagen on 4 d were significant more than those cultured in the type Ⅰ collagen on 8 d（P〈0.05）.Conclusions The characteristics of pancreatic cancer cells in two-dimensional and three-dimensional culture systems are different.MCS culture system can better mimic the in vivo growth environment of cells in tumors.