Mouse embryonic stem cells for quality control testing in assisted reproductive technology programs.
OBJECTIVE
To compare the mouse embryonic stem (ES) cell assay with the sperm motility test or 1-cell mouse embryo bioassay for embryotoxic materials.


METHODS
Cryo-preserved-thawed mouse ES-D3 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 fluorescence), 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) and 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.


CONCLUSIONS
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.
Kim
John J
JJ
Department of Gynecology and Obstetrics, Loma Linda University School of Medicine, Loma Linda, California 92354, USA. pchann@yahoo.com
Patton
William C
WC
Corselli
Johannah
J
Jacobson
John D
JD
King
Alan
A
Chan
Philip J
PJ
D000818:Animals; D017209:Apoptosis / Q000502:physiology; D001681:Biological Assay; D002470:Cell Survival; D002478:Cells, Cultured; D046149:Embryo Culture Techniques; D005260:Female; D006801:Humans; D008297:Male; D051379:Mice; D011786:Quality Control; D027724:Reproductive Techniques, Assisted / Q000592:standards*; D012680:Sensitivity and Specificity; D013081:Sperm Motility / Q000187:drug effects / Q000502:physiology*; D013234:Stem Cells / Q000166:cytology* / Q000187:drug effects