By Christine Gagliardi, Bruce A. Bunnell (auth.), Jeffrey M. Gimble, Bruce A. Bunnell (eds.)
During the earlier decade, quite a lot of clinical disciplines have followed using adipose-derived stem/stromal cells (ASCs) as an enormous device for learn and discovery. In Adipose-Derived Stem Cells: tools and Protocols, specialists from the sector, together with contributors of the esteemed overseas Federation of Adipose Therapeutics and technology (IFATS), supply outlined and verified protocols in an effort to additional codify the usage of those robust and obtainable cells. With chapters geared up round methods spanning the invention, pre-clinical, and medical approaches, a lot of the emphasis is put on human ASC, whereas extra concepts related to small and massive animal species are incorporated. As a quantity within the hugely profitable equipment in Molecular Biology™ sequence, the distinct contributions contain introductions to their respective issues, lists of the required fabrics and reagents, step by step, with ease reproducible laboratory protocols, and notes on troubleshooting and warding off recognized pitfalls. complete and state of the art, Adipose-Derived Stem Cells: equipment and Protocols serves as an essential reference textual content for knowledgeable researchers in addition to new scholars at the route to additional exploring the great capability of ASCs.
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Extra info for Adipose-Derived Stem Cells: Methods and Protocols
5) A 200-mM stock solution of insulin (2,000-fold concentration) dissolved in 10 mM HCl, made in a plastic tube, not in glass. Adipocyte maintenance medium: This solution is prepared in an identical manner as Differentiation medium, except that it does not contain either the isobutylmethylxanthine or the PPARg agonist; these two stock solutions should be omitted. Use this solution within 2 weeks of its preparation. 32 Yu et al. Freezing medium: The freezing medium consists of 80% calf serum, 10% DMEM/high glucose, and 10% dimethyl sulfoxide.
M. (2006) The immunophenotype of human adipose derived cells: Temporal changes in stromal- and stem cell-associated markers. Stem Cells 24, 376–85. Kang, S. , Putnam, L. , Popescu, I. , and Bunnell, B. A. (2004) Neurogenesis of rhesus adipose stromal cells. J Cell Sci 117, 4289–99. , Gimble, J. , and Bunnell, B. A. (2006) Biologic properties of mesenchymal stem cells derived from bone marrow and adipose tissue. J Cell Biochem 99, 1285–97. 12. Rodbell, M. (1966) Metabolism of isolated fat cells.
Media should be changed about every 3 days. 3. Aspirate the media from the flasks and rinse the cells with PBS. Add enough trypsin to just cover the surface of the flask and incubate at 37°C for 5 min (see Note 11).
Adipose-Derived Stem Cells: Methods and Protocols by Christine Gagliardi, Bruce A. Bunnell (auth.), Jeffrey M. Gimble, Bruce A. Bunnell (eds.)