Human genome-specific real-time PCR method for sensitive detection and reproducible quantitation of human cells in mice
P Song, Z Xie, L Guo, C Wang, W Xie, Y Wu - Stem Cell Reviews and …, 2012 - Springer
P Song, Z Xie, L Guo, C Wang, W Xie, Y Wu
Stem Cell Reviews and Reports, 2012•SpringerXenotransplantation of human cells into immunodeficiency mice has been frequently used to
study stem cells in tissue repair and regeneration and cancer cell metastasis. However, a
sensitive and reproducible method to quantify cell engraftment lacks. Here, we developed a
Real-Time PCR-based method which facilitated consistent detection and quantification of
small amounts of human cells distributed in mouse organs after infusion. The principle of the
method was to directly detect a humans-specific sequence in the human-murine genomic …
study stem cells in tissue repair and regeneration and cancer cell metastasis. However, a
sensitive and reproducible method to quantify cell engraftment lacks. Here, we developed a
Real-Time PCR-based method which facilitated consistent detection and quantification of
small amounts of human cells distributed in mouse organs after infusion. The principle of the
method was to directly detect a humans-specific sequence in the human-murine genomic …
Abstract
Xenotransplantation of human cells into immunodeficiency mice has been frequently used to study stem cells in tissue repair and regeneration and cancer cell metastasis. However, a sensitive and reproducible method to quantify cell engraftment lacks. Here, we developed a Real-Time PCR-based method which facilitated consistent detection and quantification of small amounts of human cells distributed in mouse organs after infusion. The principle of the method was to directly detect a humans-specific sequence in the human-murine genomic DNA mixture. In a mouse myocardial infarction model, the Real-Time PCR-based method consistently determined the amounts of human mesenchymal stem cells (hMSCs) engrafted into the heart and other organs 7 days after infusion of as little as 2.5 × 105 cells, indicating a high sensitivity, and the amounts of hMSCs detected in mice highly correlated to the numbers of hMSCs transplanted. Importantly, different from previous PCR-based methods, our method produced highly consistent and reproducible results. The reliability of the method was further proven by parallel analyses of DiI-labeled hMSCs in tissue sections and in single cell suspensions of mice. Our data show that the present human genomic DNA-specific primers-based Real-Time PCR method is sensitive and highly reproducible in determining the amount of xenotransplanted human cells in murine tissues.
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