@article {1577, title = {Quantification of Neurotransmitters from Intact and Regenerating Planarians Using UHPLC-MS/SRM Method.}, journal = {Methods Mol Biol}, volume = {1774}, year = {2018}, month = {2018}, pages = {555-570}, abstract = {

Freshwater planarian species S. mediterranea is an emerging stem cell model because of its capability of regenerating large portions of missing body parts. It is one of the best model systems available to address the basic biological mechanisms in the regeneration processes. Absolute quantification of metabolites from planarians is imperative to understand their role in the regeneration processes. Here we describe a stable isotope dilution ultrahigh performance liquid chromatography/mass spectrometry/selected reaction monitoring (UHPLC-MS/SRM) assay for a sensitive and quantitative assessment of neurotransmitters (NTs) in planaria. We used this method for the simultaneous quantification of 16 NTs from both intact and regenerating planarians.

}, keywords = {Animals, Chromatography, High Pressure Liquid, Neurotransmitter Agents, Planarians, Regeneration, Stem Cells, Tandem Mass Spectrometry}, issn = {1940-6029}, doi = {10.1007/978-1-4939-7802-1_25}, author = {Rangiah, Kannan and Palakodeti, Dasaradhi} } @article {1211, title = {Mimicking Muscle Stem Cell Quiescence in Culture: Methods for Synchronization in Reversible Arrest.}, journal = {Methods Mol Biol}, volume = {1556}, year = {2017}, month = {2017}, pages = {283-302}, abstract = {

Growing evidence supports the view that in adult stem cells, the defining stem cell features of potency and self-renewal are associated with the quiescent state. Thus, uncovering the molecular logic of this reversibly arrested state underlies not only a fundamental understanding of adult tissue dynamics but also hopes for therapeutic regeneration and rejuvenation of damaged or aging tissue. A key question concerns how adult stem cells use quiescence to establish or reinforce the property of self-renewal. Since self-renewal is largely studied by assays that measure proliferation, the concept of self-renewal programs imposed during non-proliferating conditions is counterintuitive. However, there is increasing evidence generated by deconstructing the quiescent state that highlights how programs characteristic of this particular cell cycle exit may enhance stem cell capabilities, through both cell-intrinsic and extrinsic programs.Toward this end, culture models that recapitulate key aspects of stem cell quiescence are useful for molecular analysis to explore attributes and regulation of the quiescent state. In this chapter, we review the different methods used to generate homogeneous populations of quiescent muscle cells, largely by manipulating culture conditions that feed into core signaling programs that regulate the cell cycle. We also provide detailed protocols developed or refined in our lab over the past two decades.

}, keywords = {Actins, Adult Stem Cells, Animals, Biomarkers, Cell Culture Techniques, Cell Differentiation, Cell Line, Cell Proliferation, Fluorescent Antibody Technique, Humans, Mice, Microscopy, Fluorescence, Muscle, Skeletal, Myoblasts, Resting Phase, Cell Cycle, Satellite Cells, Skeletal Muscle, Stem Cells}, issn = {1940-6029}, doi = {10.1007/978-1-4939-6771-1_15}, author = {Arora, Reety and Rumman, Mohammed and Venugopal, Nisha and Gala, Hardik and Dhawan, Jyotsna} }