Separation of rabbit red cells by density in a bovine serum albumin gradient and correlation of red cell density with cell age after in vivo labeling with 59Fe
✍ Scribed by Charles Bishop; Theodore C. Prentice
- Publisher
- John Wiley and Sons
- Year
- 1966
- Tongue
- English
- Weight
- 871 KB
- Volume
- 67
- Category
- Article
- ISSN
- 0021-9541
No coin nor oath required. For personal study only.
✦ Synopsis
Rabbit red cells are separated by centrifuging them for one hour at 33,000 G in density gradient tubes of bovine serum albumin. The separation represented an equilibrium situation since rebanding experiments showed that the cells from a layer would again seek that density layer when recentrifuged in a new gradient tube. When rabbits were injected with 59Fe, the radioactive red cells at one day were nearly all light, but these labeled cells moved into the more dense layers over the next few days. This not only shows that the separation by density is discriminating but that some red cells became dense very quickly. Bearing in mind the problems of interpreting radioactive iron data because of the possibility of reutilization, it is tentatively concluded that dense red cells are not necessarily senescent red cells since these dense cells appear to persist for the normal life span of the rabbit red cell.
As red cells mature and age, they become more dense. The separation of younger and older red cells according to their density can be achieved by centrifugation (or possibly by gravity alone) either in media that are less dense than the red cells or in media that are similar in density to the red cells. The former approach is simpler since the cells can be centrifuged in their own plasma or in saline or buffer solutions. The technic has been utilized frequently (Beard and Finulli,