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Temperature variable and the efficiency of sperm mediated transfection of HPV16 DNA into cells

Asian Journal of Andrology 2002年第3期4卷 169-173页

作者： ruslana kadze Philip J.Chan John D.Jacobson Johannah U.Corselli Alan King Center for Fertility and In Vitro Fertilization Department of Gynecology and Obstetricsand Department of Physiology and PharmacologyLoma Linda University School of MedicineLoma LindaCalifornia 92350USA

Aim:To pretreat sperm at various temperatures before exposure to human papillomavirus(HPV)16 DNA fragments and to assess the efficiency of HPV carrier sperm to transfect cumulus ***:Cumulus cells from follicular aspirates were obtained,pooled and divided into culture dishes containing Sybr Gold-stained HPV DNA carrying sperm that were either pretreated at 4℃,37℃or 40℃(n=5).The cells were incubated in 5%CO_(2) in air mixture at 37℃for 24 *** efficiency of sperm to take up fluorescent HPV DNA was determined at hour *** incubation,cumulus cell viability was assessed using the e0sin method and the percentages of fluorescent cumulus cells ***:Over half of all the cumulus cells became fluorescent with the highest percentage in the 37℃*** pretreated at 4℃had the greatest amount of HPV DNA *** sperm motility was similar for the 3 pretreatment *** were no differences in cumulus viability among the ***:Sperm pretreated at 37℃transferred the greatest amount of fluorescent HPV DNA fragments to the cumulus *** HPV DNA was observed in the nuclear and cytoplasmic *** data suggested the possibility of sperm as a vector for the transmission of HPV DNA to the cumulus cells surrounding ovulated oocytes,which might lead to early implantation failures.

关键词： spermatozoa human papillomavirus gene transfer cumulus fertility

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