Volume 21, Issue 4 (Iranian South Medical Journal 2018)
Iran South Med J 2018, 21(4): 304-318 |
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1- Department of Hematology and Blood Banking, School of Medical Sciences, Tarbiat Modarres University, Teh-ran, Iran
2- Department of Hematology and Blood Banking, School of Medical Sciences, Tarbiat Modarres University, Teh-ran, Iran , kavianis@modares.ac.ir
2- Department of Hematology and Blood Banking, School of Medical Sciences, Tarbiat Modarres University, Teh-ran, Iran , kavianis@modares.ac.ir
Abstract: (3908 Views)
Background: Multiple myeloma is a type of plasma cell carcinoma, whose development depends on the interaction between malignant plasma cells and the microenvironment that is regulated by the receptors. Although the initial adhesion of the cells, resistance to apoptosis, and increased secretion of cytokines and growth factors contribute to the survival, growth and proliferation of myeloma cells, maintaining lung cells in the laboratory is not easy and faces certain challenges. Since conducting research on myeloma requires optimizing and maintaining their culture conditions in vitro, it appears that amniotic fluid can promote the culture of these cells in vitro.
Materials and Methods: Myeloma cells were isolated from six patients by MACS technique. Cells were cultured in media containing RPMI with 10% FBS (active and inactive) and 10%, 25%, 50% amniotic fluid (active and passive) for 2 weeks. Cell survival was measured every other day using trypan blue staining during these two weeks. Next, the expression of proliferation genes (BCL-2), implantation genes (CXCR4) and cell cycle stop genes (P21, P27) were studied using qualitative PCR technique.
Results: The results showed that the medium containing 25% passive amniotic fluid and 10% inactive FBS significantly increased the proliferation of myeloma cells, and on day zero (on the day of isolation) all of these genes were expressed. Furthermore, on the fourth day of cultivation, in all groups, BCL-2 and CXCR4 genes were expressed, while P21 and P27 genes were not. This difference could indicate the effect of amniotic fluid on the growth and proliferation of the myeloma cells.
Conclusion: According to the results, the use of a medium containing 25% inactive amniotic fluid plus a base medium that contains active RPMI and FBS 10% is recommended for the culture of myeloma cells.
Materials and Methods: Myeloma cells were isolated from six patients by MACS technique. Cells were cultured in media containing RPMI with 10% FBS (active and inactive) and 10%, 25%, 50% amniotic fluid (active and passive) for 2 weeks. Cell survival was measured every other day using trypan blue staining during these two weeks. Next, the expression of proliferation genes (BCL-2), implantation genes (CXCR4) and cell cycle stop genes (P21, P27) were studied using qualitative PCR technique.
Results: The results showed that the medium containing 25% passive amniotic fluid and 10% inactive FBS significantly increased the proliferation of myeloma cells, and on day zero (on the day of isolation) all of these genes were expressed. Furthermore, on the fourth day of cultivation, in all groups, BCL-2 and CXCR4 genes were expressed, while P21 and P27 genes were not. This difference could indicate the effect of amniotic fluid on the growth and proliferation of the myeloma cells.
Conclusion: According to the results, the use of a medium containing 25% inactive amniotic fluid plus a base medium that contains active RPMI and FBS 10% is recommended for the culture of myeloma cells.
Type of Study: Original |
Subject:
Hemic and Lymphatic Systems
Received: 2016/08/30 | Accepted: 2018/02/14 | Published: 2018/09/4
Received: 2016/08/30 | Accepted: 2018/02/14 | Published: 2018/09/4
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