ABSTRACT
Background: Bone marrow mesenchymal stem cells [BM-MSCs] have emerged as a potential therapy for various inflammatory diseases. Because of some limitations, several recent studies have suggested the use of embryonic stem cell-derived MSCs [ESC-MSCs] as an alternative for BM-MSCs. Some of the therapeutic effects of the ESCMSCs are related to the secretion of a broad array of cytokines and growth factors, known as secretome.Harnessing this secretome for therapeutic applications requires the optimization of production of secretary molecules. It has been shown that aggregation of MSCs into 3D spheroids, as a preconditioning strategy, can enhance immunomodulatory potential of such cells. In this study, we investigated the effect of secretome derived from human ESC-MSCs [hESC-MSCs] spheroids on secretion of IL-1Beta, IL-10, and tumor necrosis factor Alpha [TNF-Alpha] from lipopolysaccharide [LPS]-induced peripheral blood mononuclear cells [PBMCs]
Methods: In the present study, after immunophenotyping and considering mesodermal differentiation of hESC-MSCs, the cells were nonadherently grown to prepare 3D aggregates, and then conditioned medium or secretome was extracted from the cultures. Afterwards, the anti-inflammatory effects of the secretome were assessed in an in vitro model of inflammation
Results: Results from this study showed that aggregate-prepared secretome from hESC-MSCs was able to significantly decrease the secretion of TNF-Alpha [301.7 +/- 5.906, p < 0.0001] and IL-1 Alpha [485.2 +/- 48.38, p < 0.001] from LPS-induced PBMCs as the indicators of inflammation, in comparison with adherent culture-prepared secretome [TNF-Alpha : 166.6 +/- 8.04, IL-1Beta: 125.2 +/- 2.73]
Conclusion: Our study indicated that cell aggregation can be an appropriate strategy to increase immunomodulatory characteristics of hESC-MSCs
ABSTRACT
Background: Arsenic trioxide [ATO] has been reported as an effective anti-cancer and a US Food and Drug Administration [FDA] approved drug for treatment of some cancers. The aim of this study was to determine the underlying apoptosis molecular and cellular mechanisms of ATO in the presence or absence of ionizing radiation [IR] in vitro in the glioblastoma multiforme [GBM] cell line, U87MG
Methods: Cells were treated by different concentrations of ATO either in presence or absence of IR. Viability and apoptosis pathway of both treated and control groups were evaluated using MTT assay and the expression analysis of Box, Bcl-2, and caspase-3 genes, respectively. All treatments were performed on 100-ujm diameter spheroids
Results: Results showed a significant reduction in the survival of the cells in all treated groups. As expected, cell survival was much less in combination treatment than treatment with only ATO. Moreover, combination therapy made Box and caspase-3 up-regulated and Bcl-2 down-regulated
Conclusion: ATO and radiation had a synergistic apoptotic effect on GBM cells by up-regulation of caspase-3 and alteration of the Bax-Bcl-2 balance; therefore, ATO may act as a potential anti-cancer agent against GBM cells through triggering the mitochondria! pathway of apoptosis
Subject(s)
Journal Article , Apoptosis/radiation effects , Arsenicals/therapeutic use , Oxides/therapeutic use , Radiation, Ionizing , In Vitro Techniques , Glioblastoma , Cell Line, TumorABSTRACT
Background: As a drug target and an antigenic agent, HIV-1 protease [HIV-1 PR] is at the center of attention for designing anti-AIDS inhibitors and diagnostic tests. In previous studies, the production of the recombinant protease has been faced with several difficulties; therefore, the aims of this study were the easy production, purification of the soluble form of protease in E. coli and investigation of its immunoreactivity
Methods: Protease coding region was isolated from the serum of an infected individual, amplified by RT-PCR and cloned into PTZ57R using TA-cloning. Protease coding frame was isolated by PCR and cloned in pET102/D. TOPO expression vector and cloned protease was expressed in Escherichia coli [E. coli] BL21. Produced recombinant protein was purified by affinity Ni-NTA column and protein concentration was checked by BCA protein assay kit. Subsequently, immunoreactivity of recombinant protease [rPR] was assayed by Western blotting and ELISA
Results: Cloning of the HIV protease by TOPO cloning system in pET102/D.TOPO was confirmed with PCR and sequencing. The concentration range of purified recombinant protein was 85 to 100 micro g/ml. Immunogenicity of rPR was confirmed by Western blotting and ELISA
Conclusion: Soluble production of recombinant HIV-1 protease [HIV-1 rPR] was performed successfully. This recombinant protein disclosed 86% specificity and 90% sensitivity in immunoassay tests