Purification of Mouse Bone Marrow-Derived Stem Cells Promotes Ex Vivo Neuronal Differentiation

Abstract

The main objective of this study is to test the potential of specific populations of mouse bone marrow-derived stem cells (BMSCs) to differentiate into the neuronal cell lineage. Bone marrow of 33 mice was aspirated under general anesthesia. The collected marrows were analyzed for cell counts, compositions, and percentages of different stem cell types. We used the Midi MACS magnetic separator to purify specific populations of stem cells from the aspirated bone marrow. Cells were analyzed using flow cytometry. We assessed the presence of stem cell antigen-1 (Sca-1 + ) and prominin-1 + cells in the cellular fraction that was depleted of lineage-committed cells (lineage - ). Both purified and nonpurified cells were cultured ex vivo using specific growth media with factors that drive the cells to differentiate into the neuroglial cell types. Cells were then analyzed by flow cytometry for expression of specific neuronal markers. Our results showed that there was an increase of Sca-1 + and prominin-1 + cells in the lineage - fraction over the unpurified BM. After lineage depletion, the percentages of Sca-1 + and prominin-1 + cells increased from 4.9% and 2.6%, up to 76.1% and 59%, respectively. Unpurified mouse BM differentiated into fibroblasts, whereas Sca-1 + cells were able to generate astrocytes. Interestingly, purified prominin-1 + cells were able to generate neuronal cells. Purification of adult bone marrow-derived stem cells enhances their potentiality for differentiating into specific neuronal cell lineages.