SEPARATION OF MAST CELLS BY COIL PLANET CENTRIFUGE

通过螺旋行星离心机分离肥大细胞

• 批准号: 3779527
• 负责人: Y ITO
• 金额: --
• 依托单位: NATIONAL HEART, LUNG, AND BLOOD INSTITUTE
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/3779527
• 关键词: cell sorting; centrifugation; mast cell

Rat and mouse cells were separated with an improved nonsynchronous flow- through coil planet centrifuge (CPC). Peritoneal cells were suspended at the concentration of 2-3 x 10 7 cells/ml in conditioned RPMI 1640, supplemented with 50% heat inactivated FCS and 0.32% sodium citrate. In each separation 3ml of cell suspensions were loaded into the coiled column and elutriated at 4 degrees C. Several conditions including the centrifugal force, revolution/rotation ratio, density of separation media, flow speed, designs of both coiled column and flow tubes, etc. were examined and optimized for mast cell purification. Rat mast cells were separated at the purity of 99.1%, with an average yield of 42%. Over 90% pure mouse mast cells were harvested, despite a very low population of mast cells available in murine peritoneal cells. Purified cells were biologically intact and showed little damage.

用改进的非同步流动法分离大鼠和小鼠细胞， 直通螺旋行星离心机（CPC）。 悬浮腹膜细胞 在条件化的RPMI 1640中浓度为2-3 × 10 7个细胞/ml， 补充有50%热灭活FCS和0.32%柠檬酸钠。 在 每次分离时，将3 ml的细胞悬浮液加载到盘绕的 柱，并在4 ℃下淘洗。 几个条件，包括 离心力、公转/自转比、分离密度 介质、流速、盘管和流管的设计等。 进行了检查和优化肥大细胞纯化。 大鼠肥大细胞 分离得到纯度为99.1%的目标化合物，平均收率为42%。 收获了超过90%纯的小鼠肥大细胞，尽管非常低， 在鼠腹膜细胞中可获得的肥大细胞群。 纯化 细胞在生物学上是完整的并且显示出很少的损伤。