In this study characterization of endothelial cells differentiated from human bone marrow mesenchymal stem cells (hBMCs) was investigated in relation to their capillary network formation potential. Differentiation was performed in presence of vascular endothelial growth factor (VEGF) and insulin like growth factor-1 (IGF-1). A panel of cellular and molecular markers was used for characterization of the endothelial cells. The cells were strongly positive for von Willebrand factor (vWF) and vascular endothelial growth factor receptor 2 (VEGFR2) when measured at protein and mRNA levels. Development of endothelial cells was found to be associated with formation of typical organelles such as Weibel Palade (WP) bodies, Cavealae and pinocytic vesicles. Early vessel growth was also evidenced by showing specific junctions between the cells. The migratory and angiogenic properties of the cells were confirmed by showing capillary network formation in vitro. These results indicate that the capacity of endothelial cells differentiated from hBMSCs in formation of vascular system is consistent with molecular and structural development.
Human platelet releasate is an efficient and safe substitute for FBS in culture media used for expansion and differentiation of hBMSCs to hepatocyte.
Background: Vitamin D deficiency is related to rickets in children, and it can increase the risk of osteoporosis in adulthood. The aim of our study was to estimate the prevalence of vitamin D deficiency among healthy Iranian children and adolescents. Vitamin D levels less than 20ng/ml and between 20 and 30ng/ml was considered as vitamin D deficiency and insufficiency, respectively. Methods: Relevant observational studies evaluating the prevalence of vitamin D deficiency through 1 January 1990 to 28 Dec 2016, were searched in several electronic databases including Iran-Medex, Scientific Information Database (SID), Irandoc, PubMed and NLM Gateway (for MEDLINE), Web of Science, and Scopus with no restriction on language. Only full-text articles were used for data extraction and synthesis after considering the inclusion/exclusion criteria. Results: 11 studies included; the data of four studies of Iranian newborns were withdrawn because of their high heterogeneity. The prevalence of vitamin D deficiency in Iranian boys and girls were 35% (CI 95% 34–37) and 61% (CI 95% 60–63), respectively. The prevalence of vitamin D insufficiency in Iranian children and adolescents was 31% (CI 95% 30–31). Conclusion: It seems that the prevalence of vitamin D deficiency is very high among Iranian children and adolescents. The present findings could provide practical information for healthcare decision makers.
Background Impaired breathing during sleep, as in obstructive sleep apnea (OSA), can lead to behavior symptoms like those observed in children with attention deficit hyperactivity disorder (ADHD). Obstructive sleep apnea can be effectively treated, thus avoiding problematic pharmacotherapies associated with managing ADHD. Diagnosis of OSA relies on sleep studies as the gold standard, but in children, sleep studies are inherently difficult, cumbersome, and expensive and are not practical tools in the differential diagnosis of behavior disorders. Therefore, development of clinical laboratory tests for diagnoses of sleep apnea would change the standard of care for attention deficit syndromes. Content We review the status of potential laboratory tests for diagnosis of OSA in children with emphasis on markers linked to intermittent hypoxia and cardiovascular responses. In the context of ADHD, we focus on preliminary evidence and rationale for urocortin 3 and erythropoietin as urinary markers with physiologic relevance for diagnosis of OSA. Summary Laboratory tests that correlate with both OSA and ADHD-like syndromes would be useful to diagnose root causes of behaviors and identify a subset of children who may not need psychotropic medications. The discovery of laboratory biomarkers for OSA is evolving, but several candidates show promise and provide a segue to more focused development in laboratory diagnostics.
4093 Hematopoietic stem cell (HSC) transplantation has become a common procedure for treatment of hematopoietic malignancies and autoimmune disease. Despite significant advances in HSC transplantation, the morbidity and mortality of ablative conditioning and graft-versus-host disease (GVHD) remain limitations to application in the clinic. However, these risks can be overcome through less toxic nonmyeloablative conditioning and cell depletion strategies to remove GVHD causing-cells while retaining engraftment enhancing-tolerogeneic cells. We were the first to discover CD8+/TCR− graft facilitating cells (FC) in mouse bone marrow. The addition of as few as 30,000 FC to 10,000 HSC significantly enhances engraftment of HSC in allogeneic recipients without causing GVHD. FC also potently enhance engraftment of limiting numbers of syngeneic HSC. Human CD8+/TCR- FC comprised 1.1% ± 0.27% of total G-CSF-mobilized peripheral blood mononuclear cells (mPBMC). In the CD8+/TCR- FC, 48% of cells expressed CD3ε+, 43% were FoxP3+, 43% were CD11c+, 19% were CD19+, and 30% were HLA-DR+. Approximately 55% of FC are also CD56dim/-, and the remaining population is CD56bright. The morphology of human CD8+/TCR− FC with Wright-Giemsa staining under light microscopy suggested that the human FC population is heterogeneous. Here we evaluated if human FC enhance human HSC or progenitor homing to bone marrow of NOD/SCID/IL-2rγnull (NSG) mouse recipients. CD45+CD34+ HSC and CD8+/TCR−/CD56dim/-FC were sorted from mPBMC. NSG recipients were conditioned with 1100 cGy of total body irradiation (TBI). 24 hours after TBI, 100,000 HSC with or without 300,000 CD8+/TCR−/CD56dim/- FC were transplanted into conditioned NSG recipients. Recipients were euthanized 16 hours after transplantation. Bone marrow was harvested from femurs and tibias of recipients and plated in Colony Forming Culture (CFC) Assays. Recipients of HSC plus FC generated significantly more colony formation (colonies = 110) compared with HSC alone (colonies = 65) (P = 0.011), suggesting that CD8+/TCR−/CD56dim/- FC enhanced homing of HSC or progenitors to bone marrow. To test if human CD8+/TCR−/CD56dim/- FC facilitate engraftment of human HSC in NSG mice, 300,000 CD8+/TCR−/CD56dim/- FC were mixed with 100,000 HSC and transplanted into NSG recipient mice conditioned with 325 cGy TBI. Mice that received HSC alone served as controls. At 30 days after transplantation, PBL typing showed that 34% (10 of 29) recipients of HSC alone engrafted. In contrast, 78% of recipients (n = 23) of HSC plus CD8+/TCR−/CD56dim/- FC engrafted, and donor chimerism in PB was 1.1% ± 0.8% and 4.1% ± 1.3% (P <0.05), respectively. At 6 months after transplantation, NSG recipients of HSC + CD8+/TCR−CD56dim/- FC exhibited persistent donor chimerism in PB (9.1% ± 6% vs. 3.8% ± 3.5%) (P <0.05) and significantly higher levels of donor chimerism in spleen (26.3% ± 11.8% vs. 12.3% ± 9.8%) (P <0.05) and BM (11.6% ± 4.8% vs. 2.9% ± 1.3%) (P <0.05) compared to recipients of HSC alone. Our data indicate that CD8+/TCR−/CD56dim/- FC facilitate homing of human HSC or progenitors and enhance engraftment of human HSC in NSG recipient mice. Disclosures: Bozulic: Regenerex, LLC: Employment. Ildstad:Regenerex, LLC: Equity Ownership.
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