OPCs Differentiation Assay

Creative Bioarray offers an assay to assess the differentiation of OPCs (Oligodendrocyte Progenitor Cells) into mature oligodendrocytes.

Here's an optimized general outline for an OPC differentiation assay:

1. Cell Culture: OPCs are initially cultured in vitro. These cells can differentiate into mature oligodendrocytes.
2. Stimulus for Differentiation: Various stimuli or differentiation-inducing factors may be introduced. These could include growth factors, specific signaling molecules, or other compounds known to promote OPC differentiation.
3. Differentiation Markers: Over time, the cells will begin to express markers indicative of oligodendrocyte differentiation. These might include proteins like O4, O1, and myelin basic protein (MBP), which are associated with mature oligodendrocytes.
4. Immunocytochemistry or Flow Cytometry: Immunocytochemistry or flow cytometry techniques can be used to visualize and quantify the expression of differentiation markers. This helps confirm the transition of OPCs into mature oligodendrocytes.
5. Morphological Changes: Morphological changes can also be observed under a microscope. As OPCs differentiate into oligodendrocytes, they develop a characteristic morphology with multiple processes.
6. Data Analysis: Data collected from the assay, including quantitative measurements and images, can be analyzed to draw conclusions about the efficiency and effectiveness of OPC differentiation under various conditions.

Figure 1. Oligodendrocyte differentiation model and demonstration of possible evaluations. (B) Using this model of OL culture is possible to evaluate changes in OL differentiation stages. Graph bars represent the quantification of immature OL (NG2+ staining, red) and mature OL (MBP+ staining, green) from vehicle- and UCB-treated cells at 18 DIV. (C) In this model it is also possible to evaluate changes in OL morphology through the division of mature OL in different stages of morphologic maturity like: (1) OL with poorly branched processes, (2) OL with complex branched processes and (3) OL that partially form membranes. Graph bars represent the quantification of OL in these different stages after 18 DIV in the presence or absence of UCB. (D) As demonstrated in this type of culture it is also possible to isolate a single OL in order to perform Sholl analysis and evaluate changes in OL process branching in vehicle- and UCB-treated cells.^[1]

Assessment of OPCs Differentiation

• Differentiation markers detection
• Morphological analysis

Reference:

1. Barateiro A, Fernandes A. Temporal oligodendrocyte lineage progression: in vitro models of proliferation, differentiation and myelination. Biochim Biophys Acta. 2014;1843(9):1917-1929. doi:10.1016/j.bbamcr.2014.04.018.