Development and Validation of a qPCR Analysis of Male Porcine Cell Biodistribution in Female Pigs

• Published in: Molecular therapy Vol. 20; no. 9; pp. 62 - 63
• Main Authors: Li, Maoxiang, Hill, Emily, Grover, Jeffrey, Steenwinckel, Valerie, Wolfe, Mark, Johnson, Mark, Ma, Haiyan
• Format: Journal Article
• Language: English
• Published: 01.09.2012
• ISSN: 1525-0016; 1525-0024

Stem cells, whether gene-modified or not, present a number of safety concerns after administration into human subjects. Major questions have been raised as to where stem cells migrate to after administration, whether the cells engraft and differentiate, and what their long-term fate is. Therefore, GLP-compliant preclinical biodistribution and persistence animal studies are recommended by the United States Food and Drug Administration (FDA) to evaluate the safety of potential stem cell-based products. In this study, the specificity of the TaqMan probe/primers to the male porcine sexdetermining region Y (SRY) gene sequence was determined by carrying out real-time PCR on genomic DNA isolated from male porcine MSC in the absence and presence of female pig matrix genomic DNA (gDNA). For each assay, two sets of quality control (QC) samples were prepared independently and each set included three concentrations of male pig MSC DNA (high, middle and low) in triplicate. The assay specificity, linear range, intra- and inter-assay precision, accuracy and reproducibility, and acceptance criteria are reported.