A simple PCR‐based strategy for estimating species‐specific contributions in chimeras and xenografts

Abstract

Various tissue‐engineering strategies for repair and regeneration involve using cells from different species in chimeras or xenografts. Yet a major challenge is distinguishing between donor‐versus host‐effects on gene expression. This study provides a simple molecular strategy to quantify species‐specific contributions in chimeras and xenografts using species‐specific primers for ribosomal protein L19 (RPL19). cDNA from quail, duck, chicken, mouse, and human was mixed in pairs as a dilution series, species‐specific RPL19 expression was measured, a linear regression was calculated, and the ratios of donor and host cells were estimated. To test the accuracy of the approach, quail cells were transplanted into transgenic‐GFP chick and resulting chimeras were analyzed with species‐specific primers. Fluorescent‐activated cell sorting validated the RPL19 strategy. To apply the RPL19 strategy, Runx2 expression was measured in quail‐duck chimeras. Elevated Runx2 levels correlated with higher percentages of donor cells. Finally, RPL19 primers were also shown to discriminate human‐mouse samples. Thus, this study offers a straightforward and universal strategy that enables chimeras and xenografts to be screened efficiently and rapidly prior to obtaining critical information at the molecular level. Supported by K08 DE021705 and T32 DE007306 from NIDCR to ELE.; R01 DE016402 from the NIDCR to RAS.Grant Funding Source: NIDCR