Isolation of a purified mitochondrial fraction from viable clonal insulin-producing cells (RINm5F) by percoll density gradient centrifugation |
| |
Authors: | Håkan Abrahamsson Tommy Anderson Per-Olof Berggren Håkan Pertoft |
| |
Affiliation: | 1. Department of Medical Cell Biology, Biomedicum, University of Uppsala, Sweden 2. Department of Medical Microbiology, University of Link?ping, Sweden 3. Department of Medical and Physiological Chemistry, Biomedicum, University of Uppsala, Sweden
|
| |
Abstract: | Summary A Percoll density gradient was employed for selecting large numbers of viable insulin-producing RINm5F cells. Homogenates of these cells were then subjected to gradient centrifugation and two clearly visible bands were obtained. The light fraction was essentially composed of mitochondria banded at a density of about 1.06 g/ml. The heavier fraction banded at 1.09 to 1.10 g/ml and contained lysosomes and a small number of secretory granules. The distribution of Percoll particles was restricted to the extracellular space and there was no adsorption to any membrane structures. The distribution pattern of marker enzymes for the mitochondria and lysosomes was similar to that of normal pancreatic β-cells. With the use of a Percoll density gradient it was thus possible to isolate a purified mitochondrial fraction from viable RINm5F cells. This work was supported by the Swedish Medical Research Council (03x-4, 12x-562, 12x-6240), the Swedish Diabetes Association, the Nordic Insulin Foundation, Syskonen Svenssons Foundation, and ?ke Wiberg’s Foundation. Per-Olof Berggren is a recipient of a postdoctoral fellowship from the Swedish Medical Research Council. |
| |
Keywords: | clonal insulin-producing cells (TINm5F) Percoll density gradient isolation of intact cells purified mitochondrial fraction |
本文献已被 SpringerLink 等数据库收录! |
|