Volume 26, Issue 4 (7-2019)
RJMS 2019, 26(4): 54-69 |
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Mojtahedi F, Mansouri R, Abroun S. The effect of human amniotic fluid on the survival and proliferation of human myeloma cell lines RPMI8226 and U266 in comparison with the fetal bovine serum. RJMS 2019; 26 (4) :54-69
URL: http://rjms.iums.ac.ir/article-1-5558-en.html
URL: http://rjms.iums.ac.ir/article-1-5558-en.html
, abroun@modares.ac.ir
Abstract: (3268 Views)
Background: Multiple Myeloma is a Plasma Cell Malignancy. Since the study of pathogenicity mechanisms and messenger pathways involved in the causative agent cells is important in the laboratory environment and close to the physiological environment of the body, therefore, the best environment for the study of cells in the laboratory environment, an environment most closely resembling the physiological environment of the body. However, the use of fetal bovine serum (FBS) as a supplement to the growth of cells in the cell culture process is very common. Therefore, the aim of this study was to investigate the effect of amniotic fluid on proliferation (Ki-67 and Cyclin-D) and survival (Bax and Bcl2) of both RPMI8226 and U266 Myeloma in comparison with fetal bovine serum.
Methods: Human amniotic fluid was collected from 6 pregnant women during cesarean section and sterilized. After cultivating myeloma cells (RPMI8226 and U266), cell line treatments were performed in four groups with different amniotic fluid ratios. 48 and 96 hours after treatment, cell count and cell survival were evaluated using trypan blue and neobar lam. MTT test was also used to assess the survival of the treated Myeloma cells. Finally, Ki67, Cyclin-D1, Bax and Bcl-2 genes were evaluated by Real Time PCR and SYBR Green.
Results: Evaluation of the expression of the genes involved in the process of cell proliferation and apoptosis showed that the amniotic fluid treated groups significantly increased the expression of Ki-67, Cyclin-D1 genes and also increased the anti-apoptotic gene expression of bcl 2, and a significant decrease in expression of the gene responsible for apoptosis, Bax was observed. Our data suggest that amniotic fluid as a nutrient source can increase cell growth and proliferation up to 96 hours significantly in comparison with that of fetal bovine serum, although cell proliferation and cell survival in 48 hours than 96 hours of more cultivation.
Conclusion: Based on the findings of this study, ammonia fluid can be used as a supporter of growth as a replacement for bovine serum for up to 96 hours. In this study, the effect of this fluid on the two cell lines of myeloma was evaluated, which seems to be better tested for more comprehensive conclusions in this regard on other cells.
Methods: Human amniotic fluid was collected from 6 pregnant women during cesarean section and sterilized. After cultivating myeloma cells (RPMI8226 and U266), cell line treatments were performed in four groups with different amniotic fluid ratios. 48 and 96 hours after treatment, cell count and cell survival were evaluated using trypan blue and neobar lam. MTT test was also used to assess the survival of the treated Myeloma cells. Finally, Ki67, Cyclin-D1, Bax and Bcl-2 genes were evaluated by Real Time PCR and SYBR Green.
Results: Evaluation of the expression of the genes involved in the process of cell proliferation and apoptosis showed that the amniotic fluid treated groups significantly increased the expression of Ki-67, Cyclin-D1 genes and also increased the anti-apoptotic gene expression of bcl 2, and a significant decrease in expression of the gene responsible for apoptosis, Bax was observed. Our data suggest that amniotic fluid as a nutrient source can increase cell growth and proliferation up to 96 hours significantly in comparison with that of fetal bovine serum, although cell proliferation and cell survival in 48 hours than 96 hours of more cultivation.
Conclusion: Based on the findings of this study, ammonia fluid can be used as a supporter of growth as a replacement for bovine serum for up to 96 hours. In this study, the effect of this fluid on the two cell lines of myeloma was evaluated, which seems to be better tested for more comprehensive conclusions in this regard on other cells.
Type of Study: Research |
Subject:
Immunology
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