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Type: Journal article
Title: Development of culture conditions for the isolation of pluripotent porcine embryonal outgrowths from in vitro produced and in vivo derived embryos
Author: Vassiliev, I.
Vassilieva, S.
Beebe, L.
McIlfatrick, S.
Harrison, S.
Nottle, M.
Citation: Journal of Reproduction and Development, 2010; 56(5):546-551
Publisher: Japanese Soc Animal Reproduction
Issue Date: 2010
ISSN: 0916-8818
Statement of
Ivan Vassiliev, Svetlana Vassilieva, Luke F.S. Beebe, Stephen M. Mcilfatrick, Sharon J. Harrison and Mark B. Nottle
Abstract: In the present study we examined the effect of culture media and protein source on the formation of pluripotent primary outgrowths from in vitro produced and in vivo derived porcine embryos as the first step towards the isolation of embryonic stem cells (ESCs). To do this we compared high glucose Dulbeccos Modified Eagles Medium (DMEM) with Minimal Essential Alpha Medium (αMEM) both supplemented with fetal bovine serum (FBS) or serum replacement (SR) in a 2 × 2 factorial design. Culture in DMEM or αMEM supplemented with 10% SR resulted in the establishment of homogenous populations of cells which expressed Oct 4 and Nanog. In contrast culture in either media with FBS resulted in the formation of embryonal outgrowths composed entirely of differentiated cells or a mixture of differentiated cells and putative ESCs which grew poorly and could not be passaged. Using αMEM medium containing 10% SR and culturing in 5% oxygen, putative ESC lines were isolated from in vitro and in vivo derived embryos at efficiencies of 2 and 10% respectively.
Keywords: Embryonic stem cells
Rights: (c) 2010 Society for Reproduction and Development
DOI: 10.1262/jrd.09-197A
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Appears in Collections:Aurora harvest
Obstetrics and Gynaecology publications

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