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目的观察在不同培养模式下大鼠胰腺导管来源干细胞(PDSC)的生物学特性。方法以大鼠胰腺组织为材料,采用动力性三维培养及传统二维培养方法进行细胞培养,获得原代PDSC,并连续传代,纯化PDSC,比较动力性三维培养及传统二维培养细胞间连接及细胞凋亡周期的差异。结果动力性三维培养的大鼠PDSC在培养第2天即可沿三维支架贴壁生长,在第7天即可形成细胞团,并沿三维支架多向贴壁生长;二维培养的PDSC在培养第5天贴壁生长。三维培养与二维培养相比,二维培养的细胞间连接少见,三维培养的细胞连接结构丰富。三维培养与二维培养相比,三维培养的G1期细胞增多,G2期细胞相对减少(P<0.01),三维培养早期自发凋亡细胞比例相对减少(P<0.05),晚期自发凋亡与坏死细胞比例两种培养方式间的差异无统计学意义(P>0.05)。结论动力性三维细胞培养较传统二维细胞培养,在细胞贴壁时间、细胞连接以及细胞凋亡方面具有明显的优势。
Objective To observe the biological characteristics of rat pancreatic duct derived stem cells (PDSC) in different culture modes. Methods Rat pancreatic tissues were used as materials to culture PDSCs using dynamic three-dimensional culture and traditional two-dimensional culture methods. Primary PDSCs were obtained and PDSCs were serially passaged and purified. Compared with dynamic three-dimensional culture and traditional two-dimensional culture, Differences in apoptotic cycle. Results Dynamic three-dimensional cultured rat PDSCs could grow along the three-dimensional scaffolds on the second day of culture and form cell clusters on the seventh day and adhere to the three-dimensional scaffolds in a multi-directional manner. PDSC cultured in two- Day 5 adherent growth. Three-dimensional culture compared with two-dimensional culture, two-dimensional culture of rare intercellular connections, three-dimensional culture of cell connections rich. Compared with the two-dimensional culture, the three-dimensional culture increased the number of cells in the G1 phase and the number of cells in the G2 phase (P <0.01) compared with the two-dimensional culture. The proportion of spontaneous apoptotic cells in three- There was no significant difference between the two cell culture methods (P> 0.05). Conclusion Compared with traditional two-dimensional cell culture, dynamic three-dimensional cell culture has obvious advantages in cell attachment time, cell connection and cell apoptosis.