Assessment of the purity and characteristics of rat splenic T cells isolated by one-step discontinuous gradient of percoll
dc.creator | Depamede, Sulaiman Ngongu Depamede | |
dc.date | 2012-03-04 | |
dc.date.accessioned | 2018-06-04T06:47:58Z | |
dc.date.available | 2018-06-04T06:47:58Z | |
dc.date.issued | 2012-03-04 | |
dc.description | T cells isolated from spleen lymphocytes of rat (Rattus rattus) have been commonly used in an immunological research. However the T cells are still contaminated with other splenic cells. Therefore a simple and reliable technique to isolate the T cells in one step process is needed. The aim of the present study was to assess whether T cells isolated from spleen lymphocytes of rat as a model by means of discontinuous density gradient of Percoll could give pure T cells with their activities as needed in an immunological research. Isolation was carried out in 5 density gradients of Percoll i.e. 1.052, 1.063, 1.075, 1.085, and 1.122 g/ml. The purity of the cell fractions was determined by immunocytochemistry and fluorescence-activated cell sorting (FACS) methods. The activity of each lymphocyte fraction was determined through cell proliferation assays by culturing the cells in the presence of phytohemaglutinin (PHA) and Concanavalin A (Con A) as T cells inducers. The FACS results show that fraction I and II were found to contain mostly B cells and macrophages. The fractions were obtained on the layers of gradient 1.052/1.063 and 1.063/1.075. Fraction III (gradient 1.075/1.085) contain T cells (43.2%) which consisted of CD4 (34.4%) and CD8 (18.6%), B cells (36.5%), and macrophages (10.0%). Fraction IV was strongly expressed T cell receptors (76.2%) which consisted of CD4 (63.6%) and CD 8 (20.0%), while B cells were found 20.0% and macrophages just about 0.6%. The activity of T cells as expressed by their respond to PHA and Con A shown that fraction III gave the most positive response, while fraction IV which contained the most ‘pure’ T cells gave less activity. From the present study it can be concluded that the discontinuous density gradient of Percoll method can be used to isolate and purify T cells in one step process. Furthermore, the present study indicated that T cells to proliferate in response to mitogen require other immune cells such as B cells and macrophages. Key words: Percoll, Lymphocyte, T Cell, Rat, Mitogen | en-US |
dc.format | application/pdf | |
dc.identifier | http://medpub.litbang.pertanian.go.id/index.php/jitv/article/view/653 | |
dc.identifier | 10.14334/jitv.v15i2.653 | |
dc.identifier.uri | https://repository.pertanian.go.id/handle/123456789/3118 | |
dc.language | eng | |
dc.publisher | Indonesian Animal Sciences Society | en-US |
dc.relation | http://medpub.litbang.pertanian.go.id/index.php/jitv/article/view/653/662 | |
dc.rights | Copyright (c) 1970 Indonesian Journal of Animal and Veterinary Sciences | en-US |
dc.rights | http://creativecommons.org/licenses/by/4.0 | en-US |
dc.source | 2252-696X | |
dc.source | 0853-7380 | |
dc.source | Indonesian Journal of Animal and Veterinary Sciences; Vol 15, No 2 (2010): JUNE 2010; p.157-161 | en-US |
dc.title | Assessment of the purity and characteristics of rat splenic T cells isolated by one-step discontinuous gradient of percoll | en-US |
dc.type | info:eu-repo/semantics/article | |
dc.type | info:eu-repo/semantics/publishedVersion | |
dc.type | Peer-reviewed Article | en-US |
Files
Original bundle
1 - 1 of 1
Loading...
- Name:
- Assessment of the purity and characteristics of rat splenic T cells isolated by one-step discontinuous gradient of percoll.pdf
- Size:
- 237.4 KB
- Format:
- Adobe Portable Document Format
- Description:
License bundle
1 - 1 of 1
Loading...
- Name:
- license.txt
- Size:
- 0 B
- Format:
- Item-specific license agreed upon to submission
- Description: