6 research outputs found

    Isolation of Adipose Tissue Stem Cells with Organ Culture Method

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    Background  Isolation of adipose tissue mesenchymal stem cells (MSCs) with current enzymatic methods has some limitations. For example, it is costly and time-consuming and results in a heterogeneous cell population that making compromise proliferation and differentiation of MSCs. Also, it is accompanied with the increased risk of culture contaminations because of several handling steps.In this article we present a non-enzymatic method for isolation of MSCs. Methods  Small pieces of rat adipose tissue and also human liposuction sample were placed in the culture flask, covered with fetal bovine serum (FBS) and maintained in incubator for 24 h. Then, the FBS was changed with DMEM medium containing 20% FBS. When the fibroblast-like cells were appeared around the tissue they were expanded through 3 passages and used for Immunophenotype and differentiation assays. Results  Flow cytometric analysis showed that the cells isolated with organ culture method expressed CD44 and CD105 but did not express CD34 and CD45 markers. The isolated cells also differentiated into adipocyte and osteoblast.Therefore, consistent with classically isolated MSCs, the cells isolated with our method express the stem cell surface markers and have pluripotent property. Conclusion  The presented method is an easy and cheap procedure and can be used for harvesting MSCs from very small fat samples of human or animals. Key Words: Adipocyte, Human, Osteoblast, Stem cell

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    Abstract This paper reviews recent strategies used for increasing specific yield and productivity in high cell density cultures. High cell density cultures offer an efficient means for the economical production of recombinant proteins. However, there are still some challenges associated with high cell density cultivation (HCDC) techniques. A variety of strategies in several aspects including host design consideration, tuning recombinant protein expression, medium composition, growth methodologies, and even control and analysis of the process have been successfully employed by biotechnologists to increase yield in high cell density cultures. Although most researches have focused on Escherichia coli, other microorganisms have the potential to be grown at high density and need further investigation. In recent years, information on physiological changes of hosts during different phases of cultivation derived from functional genomics, transcriptomics and proteomics is being used to overcome the obstacles encountered in high cell density cultivation and hence increase productivity

    ASA Suppresses PGE2 in Plasma and Melanocytic Nevi of Human Subjects at Increased Risk for Melanoma

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    Potential anti-inflammatory and anticarcinogenic effects of aspirin (ASA) may be suitable for melanoma chemoprevention, but defining biomarkers in relevant target tissues is prerequisite to performing randomized controlled chemoprevention trials. We conducted open-label studies with ASA in 53 human subjects with melanocytic nevi at increased risk for melanoma. In a pilot study, 12 subjects received a single dose (325 mg) of ASA; metabolites salicylate, salicylurate, and gentisic acid were detected in plasma after 4–8 h, and prostaglandin E2 (PGE2) was suppressed in both plasma and nevi for up to 24 h. Subsequently, 41 subjects received either 325 or 81 mg ASA (nonrandomized) daily for one week. ASA metabolites were consistently detected in plasma and nevi, and PGE2 levels were significantly reduced in both plasma and nevi. Subchronic ASA dosing did not affect 5” adenosine monophosphate-activated protein kinase (AMPK) activation in nevi or leukocyte subsets in peripheral blood, although metabolomic and cytokine profiling of plasma revealed significant decreases in various (non-ASA-derived) metabolites and inflammatory cytokines. In summary, short courses of daily ASA reduce plasma and nevus PGE2 and some metabolites and cytokines in plasma of human subjects at increased risk for melanoma. PGE2 may be a useful biomarker in blood and nevi for prospective melanoma chemoprevention studies with ASA
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