DNA hypomethylation of karyoplasts for bovine nuclear transplantation.

Abstract:

:The objective of this research was to evaluate if DNA hypomethylation in cells used as karyoplasts would improve development of bovine nuclear transplantation (NT) embryos. DNA from serum-fed (SF), serum-starved (SS), and 1, or 5 microM 5-azacytidine (5-aza-CR) treated cells was digested with a methylation sensitive enzyme, and evaluated for DNA methylation. A significant reduction in DNA methylation was observed in cells cultured for 48 or 72 hr in SS medium as well as in cells cultured for 48 hr in the presence of 5 microM 5-aza-CR when compared to cells cultured in SF medium. All other comparisons contained no significant differences when compared to controls. When donor cells were cultured in 5-aza-CR, SF, or SS treatment media for 48 hr, no significant difference was observed (P = 0.06) in blastocyst development rates after NT. One embryo produced by donor cells treated with 5-aza-CR established a pregnancy. Four pregnancies resulted from embryos produced by SS donor cell NT and 3 resulted from embryos produced by SF donor cell NT. Supplementation of the donor cell culture medium with 5-aza-CR was not beneficial for increasing blastocyst rate or establishing pregnancy after NT.

journal_name

Mol Reprod Dev

authors

Jones KL,Hill J,Shin TY,Lui L,Westhusin M

doi

10.1002/mrd.1079

keywords:

subject

Has Abstract

pub_date

2001-10-01 00:00:00

pages

208-13

issue

2

eissn

1040-452X

issn

1098-2795

pii

10.1002/mrd.1079

journal_volume

60

pub_type

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