Ferrer R, Planas J M, Moretó M
Rev Esp Fisiol. 1986 Sep;42(3):341-8.
The isolating agents, one enzymatic (hyaluronidase) and two chemical (sodium citrate and EDTA) have been used to search for the best technique to prepare suspensions of viable cells from chicken cecum and jejunum. Viability of enterocytes was assessed in terms of cell membrane integrity (trypan blue exclusion test), metabolic activity (oxygen uptake, lactate production and ATP content) and monosaccharide cumulative capacity. Results show that: In both cecum and jejunum, membrane integrity is better in cells harvested with citrate than those isolated with hyaluronidase or EDTA; The best metabolic status was found in cecal cells isolated with citrate and in jejunal cells obtained with hyaluronidase; The capacity to support alpha-methyl-D-glucoside gradients is highest in the cells harvested with citrate. The citrate-containing isolation medium is thus considered to yield epithelial cell suspensions with the best functional conditions.
已使用一种酶制剂（透明质酸酶）和两种化学制剂（柠檬酸钠和乙二胺四乙酸）作为分离剂，以探寻从鸡盲肠和空肠制备活细胞悬液的最佳技术。通过细胞膜完整性（台盼蓝排斥试验）、代谢活性（摄氧量、乳酸生成量和三磷酸腺苷含量）以及单糖累积能力来评估肠上皮细胞的活力。结果表明：在盲肠和空肠中，用柠檬酸盐收获的细胞的膜完整性优于用透明质酸酶或乙二胺四乙酸分离的细胞；在用柠檬酸盐分离的盲肠细胞和用透明质酸酶获得的空肠细胞中发现了最佳代谢状态；用柠檬酸盐收获的细胞中支持α-甲基-D-葡萄糖苷梯度的能力最高。因此，含柠檬酸盐的分离培养基被认为可产生具有最佳功能状态的上皮细胞悬液。
