A Nonenzymatic and Automated Closed-Cycle Process for the Isolation of Mesenchymal Stromal Cells in Drug Delivery Applications

The adipose tissue is a good source of mesenchymal stromal cells that requires minimally invasive isolation procedures. To ensure reproducibility, efficacy, and safety for clinical uses, these procedures have to be in compliant with good manufacturing practices. Techniques for harvesting and processing human adipose tissue have rapidly evolved in the last years, and Lipogems\uae represents an innovative approach to obtain microfragmented adipose tissue in a short time, without expansion and/or enzymatic treatment. The aim of this study was to assess the presence of mesenchymal stromal cells in the drain bag of the device by using a prototype Lipogems processor to wash the lipoaspirate in standardized condition. We found that, besides oil and blood residues, the drain bag contained single isolated cells easy to expand and with the typical characteristics of mesenchymal stromal cells that can be loaded with paclitaxel to use for drug-delivery application. Our findings suggest the possibility to replace the drain bag with a "cell culture chamber" obtaining a new integrated device that, without enzymatic treatment, can isolate and expand mesenchymal stromal cells in one step with high good manufacturing practices compliance. This system could be used to obtain mesenchymal stromal cells for regenerative purposes and for drug delivery

Expression of hypoxia-inducible factor 1 alpha in mononuclear phagocytes infected with Leishmania amazonensis

Increasing evidence indicates that hypoxia-inducible factor 1 alpha (HIF-1 alpha) can be upregulated indifferent cell types by nonhypoxic stimuli such as growth factors, cytokines, nitric oxide, lipopolysaccharides and a range of infectious microorganisms. In this study, the ability of the following mononuclear phagocytes to express HIF-1 alpha is reported: mouse macrophages (mM Phi)), human macrophages (W(D) and human dendritic cells (DC), parasitized in vitro with Leishmania amazonensis; as assessed by immunofluorescence microscopy. A logical explanation for HIF-1 alpha expression might be that the mononuclear phagocytes became hypoxic after L. amazonensis infection. Using the hypoxia marker pimonidazole, observation revealed that L. amazonensis-infected cells were not hypoxic. In addition, experiments using a HIF-1 alpha inhibitor, CdCl2, to treat L. amazonensis-infected macrophage cultures showed reduced parasite survival. These studies indicated that HIF-1 alpha could play a role in adaptative and immune responses of mononuclear phagocytes presenting infection by the parasite L. amazonensis. (c) 2007 Elsevier B.V. All rights reserved.114211912

Expression Of Hypoxia-inducible Factor 1α In Mononuclear Phagocytes Infected With Leishmania Amazonensis

Increasing evidence indicates that hypoxia-inducible factor 1α (HIF-1α) can be upregulated in different cell types by nonhypoxic stimuli such as growth factors, cytokines, nitric oxide, lipopolysaccharides and a range of infectious microorganisms. In this study, the ability of the following mononuclear phagocytes to express HIF-1α is reported: mouse macrophages (mMΦ), human macrophages (hMΦ) and human dendritic cells (DC), parasitized in vitro with Leishmania amazonensis; as assessed by immunofluorescence microscopy. A logical explanation for HIF-1α expression might be that the mononuclear phagocytes became hypoxic after L. amazonensis infection. Using the hypoxia marker pimonidazole, observation revealed that L. amazonensis-infected cells were not hypoxic. In addition, experiments using a HIF-1α inhibitor, CdCl2, to treat L. amazonensis-infected macrophage cultures showed reduced parasite survival. 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