Simplified PCR assay for detecting early stages of multipotent mesenchymal stromal cell differentiation

With increased demand for standardized stem cell-based assays in basic and clinical research, there is a concerted effort to develop and share quick, robust validated assays for tracking multipotent mesenchymal stromal cell (MSC) status and multipotency retention. With respect to determining differentiation capacity, classical method is to perform time-consuming histological stain assays to detect mature differentiated cell types, which can take up to 1 month or more. To accelerate identification of MSC lineage commitment, we developed and validated a simple PCR-based growth and differentiation assay to routinely detect MSC lineage commitment within 7 days. By establishing a standardized PCR assay system, critical attributes can be temporally tracked in cultured MSC. In addition to meeting the reference criteria for MSC identification, this approach is also utilized in quality testing and lot release of stem cell media products.