HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

This Article Full Text Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in Web of Science Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via Web of Science (31) Citing Articles via Google Scholar Google Scholar Articles by HAYES, E. Articles by TROUNSON, A. Search for Related Content PubMed PubMed Citation Articles by HAYES, E. Articles by TROUNSON, A.

Nuclear transfer of adult and genetically modified fetal cells of the rat

Centre For Early Human Development, Monash Institute of Reproduction and Development, Monash University, Clayton, Victoria 3168, Australia

The present study examines the handling, activation, and micromanipulation of rat eggs in an attempt to produce live young using nuclear transfer (NT) of adult and genetically modified rat fetal cells. Mature rat eggs cultured in calcium-free medium showed reduced rates (24%) of chromosomal dispersion ("spontaneous activation" characteristic of this species) compared with eggs cultured in calcium-containing medium (47%), but failed to survive micromanipulation procedures. High rates of parthenogenetic cleavage were obtained with chemical activation using ethanol/cycloheximide (65%) compared with other standard chemical activation methods (4–28%). This type of activation was also effective in reestablishing cleavage capability (19–71%), in a time-dependent manner, of spontaneously activated eggs arrested at a second prophase-like state. At most, two of four tested micromanipulation procedures were effective in producing NT embryos capable of morula or blastocyst development (14–16%) in vivo following transfer to mouse oviducts. NT blastocysts produced from cumulus cells and transfected rat fetal fibroblasts appeared morphologically and karyotypically normal (2n = 42). Nocodazole-assisted metaphase enucleation and piezoelectric-assisted donor cell injection produced significant and equivocal effects on survival and cleavage rates of reconstructed embryos but failed to significantly improve in vivo morula/blastocyst development rates (16–28%) compared with unassisted micromanipulation (16%). Live births have not yet been obtained from early cleavage stage embryos (n = 269) transferred to pseudopregnant recipient rat oviducts. Improvements in reconstituted NT embryo culture and transfer are required for these methods to be an effective means of transgenic rat production.

activation; transfection; embryo; oocyte; transgenic

This article has been cited by other articles:

				L. J. Mullins, I. Wilmut, and J. J. Mullins Nuclear transfer in rodents J. Physiol., January 1, 2004; 554(1): 4 - 12. [Abstract] [Full Text] [PDF]

				A. Krivokharchenko, E. Popova, I. Zaitseva, L. Vil'ianovich, D. Ganten, and M. Bader Development of Parthenogenetic Rat Embryos Biol Reprod, March 1, 2003; 68(3): 829 - 836. [Abstract] [Full Text] [PDF]

				X. J. Yin, T. Tani, I. Yonemura, M. Kawakami, K. Miyamoto, R. Hasegawa, Y. Kato, and Y. Tsunoda Production of Cloned Pigs from Adult Somatic Cells by Chemically Assisted Removal of Maternal Chromosomes Biol Reprod, August 1, 2002; 67(2): 442 - 446. [Abstract] [Full Text] [PDF]