In vitro and in vivo characterization of putative porcine embryonic stem cells.
We have developed a new method for the isolation of porcine embryonic stem cells (ESCs) from in vivo-derived and in vitro-produced embryos. Here we describe the isolation and characterization of several ESC lines established using this method. Cells from these lines were passaged up to 14 times, during which they were repeatedly cryopreserved. During this time, ESCs maintained their morphology and continued to express Oct 4, Nanog, and SSEA1. These cells formed embryoid bodies in suspension culture, and could be directed to differentiate into various lineages representative of all three germ layers in vitro. When injected into blastocysts these cells localized in the inner cell mass of blastocysts. To examine their pluripotency further, cells were injected into host blastocysts and transferred to recipient animals. Of the six transfers undertaken, one recipient became pregnant and gave birth to a litter of one male and three female piglets. Microsatellite analysis of DNA extracted from the tail tissue of these piglets indicated that two female piglets were chimaeric.
Vassiliev
Ivan
I
Reproductive Biotechnology Group, Discipline of Obstetrics and Gynaecology and Centre for Stem Cell Research, University of Adelaide , Adelaide 5005, Australia .
Vassilieva
Svetlana
S
Beebe
Luke F S
LF
Harrison
Sharon J
SJ
McIlfatrick
Stephen M
SM
Nottle
Mark B
MB
D000818:Animals; D001755:Blastocyst / Q000166:cytology; D018929:Cell Culture Techniques; D002454:Cell Differentiation; D002460:Cell Line; D019070:Cell Lineage; D015925:Cryopreservation; D053595:Embryonic Stem Cells / Q000166:cytology*; D005260:Female; D005838:Genotype; D008957:Models, Genetic; D010053:Ovary / Q000378:metabolism; D011247:Pregnancy; D011270:Pregnancy, Animal; D013552:Swine