Many regenerative medical products (human cell/tissue-based products) are recently under development, and non-clinical safety studies for regenerative therapies are requested.
Especially, a biodistribution assessment of drugs is the most important. Many methods, such as quantitative polymerase chain reaction (qPCR), flow cytometry (FCM) and immunohistochemistry, have been reported for detecting human cells in animals. Since each detection method has both advantages and disadvantages, an optimal assessment system is needed for each product.
The purpose of this research was to validate qPCR and FCM methods for analysing blood concentration of human mesenchymal stem cells (MSCs) in mice. Human specific Alu PCR was applied for the quantification of human MSCs in mice blood. Antibodies to human MSCs specific cell surface markers (CD73, CD90 and CD105, positive MSCs markers) were analysed for FCM. Both methods were compared on same blood samples from male immunodeficient mice administered human MSCs intravenously. Parameters (accuracy, precision, sensitivity) were evaluated for both qPCR and FCM methods. Blood samples were collected by continuous blood sampling and were divided for qPCR and FCM methods. A standard curve for measuring cell counts of human MSCs was constructed.
We will report the assay methods to detect human cells in mice by using qPCR and FCM methods.
