| Autor: |
Chitra.S, Ganesan, Nalini, T. S., Lokeswari |
| Předmět: |
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| Zdroj: |
Sri Ramachandra Journal of Medicine; Jan-Jun2014, Vol. 7 Issue 1, p2-8, 7p |
| Abstrakt: |
Monocytes developed from bone marrow enter circulation and constitute approximately 10% of peripheral blood leucocytes in humans. Under appropriate stimulatory conditions the blood monocytes are differentiated into macrophages. The human monocyte cell line (THP 1) cells are premonocytes committed to the monocytic cell lineage and are frequently used as a model system for monocytes. These cells were cultured in the presence of human serum for the induction of terminal differentiation to macrophage like cells. Macrophages are chief source of cytokines and co stimulatory molecules that play a major role in the systematic activation of T and B cells. They are involved in many inflammatory disease conditions such as rheumatoid arthritis. In the present study, a cell culture method to ascertain the stages during macrophage differentiation was examined. The main objective of the study was to isolate and compare the differentiation of monocytes to macrophages at different time points in growth media from (i) Human peripheral blood. (ii) THP1 cells (in vitro) Methods: Five ml of peripheral blood was collected in EDTA vacutainers from healthy volunteers, ranging in age from 18 to 50 years. Monocytes prepared from samples of peripheral blood and THP1 cell lines were grown with RPMI 1640, 10% FCS and human serum in vitro. Conditions needed to optimize differentiation to macrophages were determined using viability and morphology as indicators. Cell viability was assessed by haemocytometer using 0.2% Trypan blue. Morphological differences were measured by inverted phase contrast microscope. The statistical analysis was performed using MS Excel software. The cell size measurements were made by using image pro3 software. Results and Discussions: The peripheral blood mononuclear cells (PBMC) and THP1 monocytic cells were induced with human serum and morphological differences were observed in both. The population of THP 1 monocytic cell differentiaion was more than the PBMC on third day post induction. Morphological differentiation of PBMC and THP1 cells post induction continued to be seen upto 7 days of culture indicating their ability to be functional macrophages. Of the two cells cultured PBMC cells showed greater increase in size from 6 µm to 31 µm on day 7 post induction than the THP 1 cells from 16µm to 23 µm at day 7. The results of this study are useful for optimizing the time points suitable for screening molecules in signal transduction pathway during inflammation. Conclusion: This study has revealed that the 3rd day post induction with the use of AB human serum was ideal for differentiation of cells. This model could be used to study the signal transduction pathway in rheumatoid arthritis. [ABSTRACT FROM AUTHOR] |
| Databáze: |
Complementary Index |
| Externí odkaz: |
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