Mouse embryonic stem cells for quality control testing in assisted reproductive technology programs
Kim, J.J.; Patton, W.C.; Corselli, J.; Jacobson, J.D.; King, A.; Chan, P.J.
Journal of Reproductive Medicine 50(7): 533-538
2005
ISSN/ISBN: 0024-7758 PMID: 16130852 Document Number: 585526
OBJECTIVE: To compare the mouse embryonic stein (ES) cell assay with the sperm motility test or 1-ccll mouse embryo bioassay for embryotoxic materials.STUDY DESIGN: Cryopreserved-thawed mouse ESD3 cells, 1-cell mouse embryos and donor sperm were incubated for 1-4 days in culture medium exposed to a control and 4 different test materials. ES cell viability (eosin method), apoptosis (Sybr-Gold flourescence), development of blastocysts and sperm motility parameters were measured.RESULTS: The initial viabilities of ES cell were determined to be 37.0 +/- 4.2% (n=225) aiid 54.8 +/- 7.4% (n = 218) by the eosin and Sybr Gold methods, respectively. Reduced viability of ES cells in latex glove-treated medium (25.6 +/- 0.3% and 25.7 +/- 0.3% versus control, 32.8 +/- 0.2% and 33.5 +/- 1.0% by eosin or Sybr Gold, respectively, p < 0.05) was consistent with standard bioassays. However, toxicity in the syringe was detected only by the ES cell assay. The ES cell assay sensitivities were 33% and 67% (eosin and Sybr Gold methods, respectively), and specificities were 100% for both methods.CONCLUSION: Mouse ES cell assay based on Sybr-Gold asssessment was as effective as standard bioassays for detecting embryotoxicity. The results suggested that the mouse ES assay could be used for testing contact materials and DNA-modifying agents. More studies are needed to refine and enhance the sensitivity of the ES cell assay for routine use in assisted reproductive technology clinics.