Скачать или смотреть Isolation and cultivation of enriched oligodendrocytes from post-natal mouse tissues

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The process of deriving enriched oligodendrocyte cultures and establishing oligodendrocyte/neuron myelinating co-cultures from post-natal murine tissues involves several meticulous steps aimed at isolating and cultivating these critical cell types. Initially, post-natal murine brain tissues are harvested, typically from mice aged between 0 to 21 days, as this period is crucial for oligodendrocyte development. The tissues are then subjected to enzymatic digestion, often using a combination of trypsin and DNase, to dissociate the cells while preserving their viability. Following digestion, the cell suspension is filtered through a series of mesh filters to remove debris and larger aggregates, resulting in a single-cell suspension that can be plated for further culture.

Once the oligodendrocyte precursor cells (OPCs) are isolated, they are cultured in a specialized medium that promotes their growth and differentiation into mature oligodendrocytes. This medium typically contains growth factors such as platelet-derived growth factor (PDGF) and fibroblast growth factor (FGF), which are essential for the proliferation of OPCs. Over time, as the cells mature, they begin to express specific markers indicative of oligodendrocyte identity, such as Olig2 and myelin basic protein (MBP). The culture conditions are carefully monitored to ensure optimal growth, with regular media changes and the addition of necessary supplements to support cell health and differentiation.

In parallel, the establishment of oligodendrocyte/neuron myelinating co-cultures is achieved by introducing neurons into the oligodendrocyte cultures. This co-culture system allows for the investigation of the interactions between oligodendrocytes and neurons, particularly in the context of myelination. Neurons are typically derived from embryonic or post-natal murine tissues and are plated alongside the oligodendrocytes. The co-culture environment facilitates the study of myelination processes, as oligodendrocytes extend their processes to wrap around neuronal axons, forming myelin sheaths. This setup not only provides insights into the cellular mechanisms underlying myelination but also serves as a valuable model for understanding demyelinating diseases and potential therapeutic interventions.