OPC differentiation assay
Renovo’s OPC differentiation assay is used to identify compounds that promote the production of oligodendrocytes from OPCs. In this assay, OPCs are cultured with or without compounds in differentiation media in 96-well plates. Following 5 days of differentiation, cells are stained and imaged in high-content ArrayScan™ reader in multiple channels. Computer algorithms are used to quantify the number of viable and pyknotic cells, and the number of EGFP+ oligodendrocytes in each well of the plate. Immunostaining of additional markers of different cell types can also be performed and quantified.
Applications of the OPC differentiation assay include:
- Screening of a library of compounds to identify potential candidates that promote OPC differentiation.
- Secondary screening of hits to confirm and qualify compounds for further pharmacological testing.
- Determination of dose response and EC50 of lead compounds.
- Investigation of potential interactions of lead compounds with other compounds to promote OPC differentiation.
- Identification of possible involvement of a lead compound in specific OPC signaling pathways.
Differentiation of OPCs into EGFP+ oligodendrocytes: CTL, no treatment; T3, treatment with thyroid hormone T3. OPCs were derived from PLP-EGFP transgenic mice. Cells were imaged in a high-content ArrayScan reader using a 10x objective. Green, PLP-EGFP+ oligodendrocytes; blue, Hoechst-stained nuclei.
Immunofluorescence staining of oligodendrocytes with MBP antibody:Cells were imaged in a high-content ArrayScan reader using a 20x objective. Red, MBP+ oligodendrocytes; blue, Hoechst-stained nuclei.