参环毛蚓肠上皮细胞体外培养及其生物学特性

Culture and Characterization of Intestine Epithelial Cells from Pheretima aspergillum(E. Perrier) In Vitro

该研究采用混合酶消化法获得参环毛蚓(Pheretima aspergillum)肠上皮细胞(intestine epithelial cell,IEC),并进行体外培养,成功传代培养至第8代,命名为IEC-P.A.。通过光学显微镜和透射电镜技术对IEC-P.A.细胞形态、超微结构进行了鉴定,采用MTT法绘制了细胞生长曲线,应用流式细胞术检测了DNA含量并分析其细胞周期和增殖活性。体外培养的参环毛蚓肠上皮细胞汇合后呈典型的"铺路石样"形态,细胞直径为8±2μm,表面具有大量的微绒毛,胞质内含丰富的线粒体及粗面内质网。细胞周期检测结果显示,G0/G1期细胞约占总细胞数的93%。染色体核型分析结果表明,参环毛蚓肠上皮细胞为二倍体,2n=22,核型为n(♂)=x=6m+3sm+2st。该研究成功获得了离体培养的参环毛蚓肠上皮细胞及其生物学特性的相关资料。

The intestine epithelial cell（IEC）, designated IEC-P.A., was digested by complex enzyme from Pheretima aspergillum（E. Perrier）, which has been continuously subcultured 8 times in vitro. IEC-P.A. was identified by morphological and ultrastructure observation under inverted microscope and transmission electron microscope. The growth curve of IEC-P.A. was determined by MTT assay. Cell cycle and cell proliferation were observed by flow cytometry. IEC-P.A., cultured in vitro, was about 8±2 μm in diameter, and appeared some distinct typical morphological characteristics of epithelial cells, such as cobblestone-like appearance, numerous microvilli, rough endoplasmic reticulum and mitochondria. Flow cytometric test result showed that more than 93% IEC-P.A. was in G0/G1 phase. Chromosome typing of IEC-P.A. revealed that the diploid chromosome number was 2n=22, and the chromosomal karyotype was n（♂）=x=6m＋3sm＋2st. IEC-P.A. was successfully cultured in vitro and its basic information on biological characteristics was investigated in this study.