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Components:
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Lectures on Mouse MSCs
•Introduction to stem cell biology and hierarchies within the murine skeletal system
•Phenotypic and functional dissection of MSC populations in adult mouse compact bone
•Isolation of mouse MSCs from compact bone and bone marrow
•Optimization of culture conditions for in vitro mouse MSC expansion
Laboratory on Mouse MSCs
•Hands-on aseptic dissection of mouse bones
•Detailed instruction in preparation, cleaning and crushing of mouse compact bones
•Hands-on compact bone preparation and MSC isolation
•Set-up of mouse MSC cultures with primary mouse compact bone- and bone marrow-derived cells
•Identification and enumeration of mouse compact bone- and bone marrow-derived CFU-F colonies
•Analysis of adipogenic, osteogenic and chondrogenic differentiation cultures
Lectures on Human MSCs
•Characteristics of human MSCs: tissue source, frequency, phenotype, expansion and differentiation
•Culture methods and heterogeneity of human MSC subpopulations
•Enrichment methods for human MSCs
•Characteristics and functional assays of MSCs cultured in serum- and xeno-free (XF) medium
•Clinical trials: Therapeutic applications of MSCs
Laboratory on Human MSCs
•Culture of human bone marrow-derived MSCs in serum-containing vs. serum- and animal component vs. free medium with:
•Microscopic evaluation of MSC cultures
•Subculture/passaging MSCs (with cell viability, fold expansion and optimal plating density calculations)
•Staining and enumeration of CFU-F
•Staining cells for phenotype analysis by flow cytometry
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