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Optimizing qPCR method to detect human mesenchymal stem cells in rodent tissues

Understanding the biodistribution profile of cell-based therapy products in an animal model is important in evaluating its safety and efficacy. Real-time PCR method (qPCR) is useful in detecting and quantifying the cell derived DNA in rodent tissue because it is quantitative and highly sensitive. For the evaluation of human-derived cell distribution in rodents, Alu sequence has been widely used as a target sequence of the qPCR assay due to its high copy number per cell. However, there have not been enough studies to determine whether the Alu sequence is the best target for evaluating the biodistribution of human-derived cells in rodents.

Optimizing qPCR to detect human cells in mice

qPCR testing in laboratory setting

Compared to the Alu sequence, which is the most abundant short interspersed elements (SINE) in humans, qPCR assay that targets DUF1220, a SINE with lower copy numbers, showed higher sensitivity and quantification ability in detecting human cell-derived DNA in rodents. We suggest that the total copy number of the targeted sequence can be one of the important factors in determining the quality of qPCR assay to evaluate the biodistribution of human-derived cells in rodents.

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Yukiko Arimura1, Yurie Okawa1, Akiko Suzuki1, Tomoko Yoshifuku1, Yosuke Numata1, Asako Uchiyama1, Tatsuya Jikuzono1, Sakae Kohara2

1 Shin Nippon Biomedical Laboratories, Ltd. Drug Safety Research Laboratories, 2 Shin Nippon Biomedical Laboratories, Ltd. Pharmacokinetics and Bioanalysis Center