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Mammalian Expression Systems and Transfection Techniques
To delineate the function of a single ion channel subtype amongst the multitude that normally constitutes a signalling pathway, it is often insightful to study the function or signalling of that one ion channel in the absence of the others. Mammalian cell lines that do not normally express the gene of interest can be manipulated to do so via plasmid DNA expression vectors. However, large and highly charged molecules like DNA cannot passively diffuse through cell membranes. Therefore introducing nucleic acid into mammalian cells may involve introducing pores transiently into the cell membrane to allow the passage of circula...
Source: Springer protocols feed by Cell Biology - April 12, 2013 Category: Cytology Source Type: news

Approaches to Cloning of Pain-Related Ion Channel Genes
Molecular pain research is a relatively new and rapidly expanding field that represents advancement in conventional pain research. One of the fundamentals of molecular pain involves the cloning of genes and especially the ion channels specifically involved in nociceptive processing at the periphery and centrally. A variety of approaches were used to isolate these critically important genes. Cloning of these genes involved innovative strategies based on existing molecular approaches. This review will discuss well-utilized cloning approaches and their exploitation in molecular pain research. (Source: Springer protocols feed by Cell Biology)
Source: Springer protocols feed by Cell Biology - April 12, 2013 Category: Cytology Source Type: news

Methods for Assessing Effects of Wnt/β-Catenin Signaling in Senescence of Mesenchymal Stem Cells
Mesenchymal stem cells (MSCs) represent a main population of stem cells and can differentiate into multiple cell lineages. Recently, MSC transplantation has been applied to repair the malfunctioned tissues. However, increasing evidences show that some MSCs expanded in vitro and in the aged individuals become senescent. Capacity of senescent MSCs in repairing the tissues may decrease significantly. Interestingly, preventing MSC senescence is a powerful potential strategy to delay aging of individuals and promote application of cell therapy for treating aging-related diseases. Therefore, it is necessary to explore mechanisms...
Source: Springer protocols feed by Cell Biology - February 13, 2013 Category: Cytology Source Type: news

Isolation of Mesenchymal Stem Cells from Human Bone and Long-Term Cultivation Under Physiologic Oxygen Conditions
Bone-derived stroma cells contain a rare subpopulation, which exhibits enhanced stemness characteristics. Therefore, this particular cell type is often attributed the mesenchymal stem cell (MSC). Due to their high proliferation potential, multipotential differentiation capacity, and immunosuppressive properties, MSCs are now widely appreciated for cell therapeutic applications in a multitude of clinical aspects. In line with this, maintenance of MSC stemness during isolation and culture expansion is considered pivot. Here, we provide step-by-step protocols which allow selection for, and in vitro propagation of high quality...
Source: Springer protocols feed by Cell Biology - February 13, 2013 Category: Cytology Source Type: news

Cardiac Stem Cell Senescence
Cellular senescence processes affecting tissue resident stem cells are considered, at present, an hallmark of both aging and age-related pathologies. Therefore it is mandatory to address this problem with adequate techniques that could highlight the molecular alterations associated with this complex cellular response to stressors. Here we describe methods to characterize cardiac stem cell (CSC) senescence from a molecular and functional standpoint. (Source: Springer protocols feed by Cell Biology)
Source: Springer protocols feed by Cell Biology - February 13, 2013 Category: Cytology Source Type: news

Human Myoblasts from Skeletal Muscle Biopsies: In Vitro Culture Preparations for Morphological and Cytochemical Analyses at Light and Electron Microscopy
We describe protocols for the isolation of satellite cells from human muscle biopsies, for the in vitro culture of proliferating and differentiating myoblasts, and for the preparation of cell samples suitable for morphological and cytochemical analyses at light and electron microscopy. The procedures described are especially appropriate for processing small muscle biopsies, and allow obtaining myoblast/myotube monolayers on glass coverslips, thus preserving good cell morphology and immunoreactivity for protein markers of myoblast proliferation, differentiation, and senescence. (Source: Springer protocols feed by Cell Biology)
Source: Springer protocols feed by Cell Biology - February 13, 2013 Category: Cytology Source Type: news