Azadeh Haghighitalab; Maryam M. Matin; Fatemeh Khakrah; Ahmad Asoodeh; Ahmad Reza Bahrami
Abstract
Despite the prominent therapeutic potentials of stem cells, their use in cell therapy has been challenged with some unreproducible and inconsistent outcomes in addition to the risk of rejection and tumorigenesis. Gaining novel insights to the importance of the conditioned medium, ...
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Despite the prominent therapeutic potentials of stem cells, their use in cell therapy has been challenged with some unreproducible and inconsistent outcomes in addition to the risk of rejection and tumorigenesis. Gaining novel insights to the importance of the conditioned medium, secretory factors and extracellular vesicles as the functional components of the cultured stem cells, suggested the idea of substituting the cells with their cell-free counterparts. Biological properties of these products are influenced by the cues received from their microenvironment. Hence, providing optimal and fully defined culture conditions is essential for their preparation. Fetal bovine serum (FBS), one of the most routine supplements of cell culture, is enriched by endogenous extracellular vesicles (EVs). These EVs will affect the yield, purity and functional features of the cell-free products. Here, we endeavored to examine and compare three different methods including ultrasonication, ultrafiltration and polymer-based precipitation, to deplete EVs from FBS. We chose easy to perform and fast methods with the capacity for high-throughput applications. Based on our observations, although all examined methods were able to deplete EVs from FBS to some extent, polymer-based precipitation could be considered as the method of choice with minimal consequences on the biological requirements of FBS to support cell growth and characteristics. Due to similarities between FBS and some other biological solutions, this strategy would be suitable for EV-depletion from other liquids with high concentrations of proteins and nutrients. Moreover, it could be applied for preparation of optimal culture conditions for nanoparticle applications.
Samaneh Sepahi; Razieh Jalal; Behnaz Toluinia; Ahamad Asoodeh; Jamshid Darvish
Abstract
The prevalence of type 2 diabetes mellitus (T2DM) is rising dramatically in the Middle East, especially in the Islamic Republic of Iran, but the genetic basis of type 2 diabetes in Iran is poorly understood. Polymorphisms of hepatocyte nuclear factor-1α (HNF-1α) and glucagon-like peptide-1 receptor ...
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The prevalence of type 2 diabetes mellitus (T2DM) is rising dramatically in the Middle East, especially in the Islamic Republic of Iran, but the genetic basis of type 2 diabetes in Iran is poorly understood. Polymorphisms of hepatocyte nuclear factor-1α (HNF-1α) and glucagon-like peptide-1 receptor (GLP-1R) genes showed association with type 2 diabetes in several ethnic groups. In this study, we evaluated whether these markers confer susceptibility to T2DM in a diabetic population living in Mashhad (northeast of Iran). Genotyping of Ala98Val (HNF-1α) and Thr149Met (GLP-1R) was done by the restriction fragment length polymorphism-PCR (RFLP-PCR) method in the following groups: 1) early-onset diabetes (age at onset ≤ 35 years); 2) late-onset diabetes (age at onset > 35 years); and 3) control. Our results showed that CT (Ala/Val) genotype of HNF-1α was higher in the early-onset type 2 diabetic group compared to the controls but difference was not significant. We did not find the GLP-1R Thr149Met mutation in all participants. The prevalence of the HNF-1α (Ala98Val) and (GLP-1R) Thr149Met mutations has not been previously reported in Iranian participants. We conclude that these mutations are not a common cause of T2DM in our studied population.
