Stem Cell Research
Sample Data
Over the last decade, interest has rapidly grown beyond hematopoietic stem cells to understand the role of embryonic and other adult (or somatic) stem cell populations. BD flow cytometry analyzers can be used in combination with fluorochrome-conjugated antibodies to count members of cell subpopulations and obtain data on the relative expression level of multiple markers for individual self-renewing or differentiated cells.
Mesenchymal stem cells grown under serum and serum-free conditions
In the experiment shown, scientists at BD Technologies used integrated propriety bioinformatics tools and automation to conduct an advanced combinatorial screen of large numbers of biological factors to develop a custom, serum-free culture environment for mesenchymal stem cells (MSCs). MSCs were seeded onto 6-well plates in serum-free medium with an optimized growth-factor combination on a selected extracellular matrix surface. Serum-containing cultures were grown on tissue culture polystyrene (TCPS). At confluency (day 5), all cells were removed, and cell number was counted on a hemocytometer. Cells were then replated under the same conditions for another passage. Cell number was counted again at confluency (day 7). The serum-free environment greatly outperformed the serum-containing control over both passages. Serum-free growth on extracellular matrices gave 5.4 and 6.5-fold expansion vs. 3.5 and 3.1 fold in serum on TCPS, respectively. These results demonstrate that this serum-free culture system provides a better environment for expansion, but also allows the cells to maintain phenotype, thus retaining the ability to differentiate down multiple lineages.
Mesenchymal stem cells induced to differentiate into osteocytes
For many applications, cell-based assays require extensive, potentially time-consuming optimization to define the optimum components necessary to monitor expansion or differentiation down a specific lineage. In this experiment, scientists using BD Technologies Custom Discovery Platform developed an optimized assay to confirm differentiation of mesenchymalstem cells into osteocytes. The assay was tested using the BD Pathway™ 855 bioimaging system. Cells were stained with Alexa Fluor® 594 and Hoechst nuclear DNA stain.
