The effect of amniotic membrane on growth, proliferation, and survival of the myeloma cells and examination of genes related to proliferation (BCL2), implantation (CXCR4), and cell cycle stop (P21 and P27)

Abstract

BACKGROUND: The myeloma cell is not able to grow and proliferate out of bone marrow (BM) media, and in laboratory conditions its survival is low. We considered to use an environment that has the same conditions as body physiological conditions. In this study, the effect of the amniotic membrane (AM) on the growth and proliferation of myeloma cells were evaluated. METHODS: This study was performed on plasma cells derived from BM aspiration (primary cells) in 3 patients with multiple myeloma (MM). Plasma cells of these patients were isolated by magnetic-activated cell sorting (MACS) technique and cultured in different environments of AM for two consecutive weeks, and then were examined by qualitative polymerase chain reaction (PCR) technique for expression of genes related to proliferation [B-cell lymphoma 2 (BCL2)], implantation [chemokine receptor type 4 (CXCR4)], and cell cycle stop (P21, P27). RESULTS: Isolated plasma cells were cultured in 3 different groups for 2 weeks. The most cell proliferation was observed in the medium containing Roswell Park Memorial Institute (RPMI) medium from amniotic cultures and plasma cells [an environment without fetal bovine serum (FBS)]. All genes were expressed on day zero (on the day of isolation). On the day 4, proliferation genes (BCL2) and implantation genes (CXCR4) had an expression in the control group without FBS medium, but P21 and P27 genes had no expression. CONCLUSION: The best environment for the growth and maintenance of plasma cells in vitro is the use of RPMI from the AM (without FBS) in which plasma cells can be kept alive for 10 days.