兔心房来源非心肌细胞的分离培养

Isolation and cultivation of nonmyocytes derived from the atrial wall in rabbit

建立一种兔心房组织来源的非心肌细胞体外培养模型,为心血管药理实验、病毒学实验及组织工程的研究提供实验平台.无菌条件下取兔心房壁组织,将组织分离成2 mm×1 mm小块,采用植块法以5行4列的形式种植于培养瓶中,加入M199或MEM培养基对兔心房组织进行原代培养.待长成单层时胰酶消化收集细胞,并进行传代培养和细胞冻存.该方法获得了大量活力良好的兔心房壁组织来源的非心肌细胞,倒置镜下观察细胞多呈短梭形,以及少量的三角形、柱形和卵圆形细胞,细胞核部位隆突,胞浆均匀清亮.经体外传代培养至16代,冷冻复苏以后,细胞仍生长良好.

The study established the model of nonmyocytes derived from the atrial wall of rabbit in vitro, and provided a platform for cardiovascular pharmacological experiments, virology experiments and the research of tissue engineering. For the primary culture, the atrial wall of rabbit was isolated in the sterile condition, then was dissociated into 2 mm×1 mm little parts that eventually paved on the bottom of cultivation flask by 5 rows and 4 columns with M199 or MEM medium, immerged its base. When cells grew into the monolayer, trypsinized and harvested a great deal of energetic cells as well as the subculture and cell freezing were performed. Under the convert phase microscope, nonmyocytes derived from the atrial wall presented a short shuttle with the prominency nucleus and homogeneous transparent cytoplasm, together with little bit other cells, such as triangle cells, column-like cells and ovum-like cells. Nonmyocytes subcultured to passage 16 in vitro and the cryopreservation was carried out, and cells grew still vigorously after defreezing.