Establishment of pluripotent cell lines from porcine preimplantation embryos

Embryonic stem (ES) cells are pluripotent cells isolated from in vitro culture of preimplantation embryos. Experiments were undertaken to identify preimplantation embryonic stages and culture conditions under which pluripotent, porcine embryo-derived cell lines could be isolated. Cell lines were est...

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Bibliographic Details
Published in:Theriogenology 1999-07, Vol.52 (2), p.195-212
Main Authors: Chen, L.R., Shiue, Y.L., Bertolini, L., Medrano, J.F., BonDurant, R.H., Anderson, G.B.
Format: Article
Language:English
Subjects:
alkaline phosphatase
Alkaline Phosphatase - analysis
Animals
blastocyst
Blastocyst - cytology
Cell Culture Techniques - methods
Cell Differentiation
Cell Division
cell lines
Cells, Cultured
chimerism
embryo (animal)
embryo culture
Embryo, Mammalian
embryogenesis
embryonic stem cells
enzyme activity
Female
Fibroblasts - cytology
Karyotyping
Mice
nuclear transfer embryos
nuclei
Oocytes - cytology
pig
porcine
preimplantation period
Stem Cells - cytology
Swine
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Summary:Embryonic stem (ES) cells are pluripotent cells isolated from in vitro culture of preimplantation embryos. Experiments were undertaken to identify preimplantation embryonic stages and culture conditions under which pluripotent, porcine embryo-derived cell lines could be isolated. Cell lines were established from in vitro culture of intact, porcine early hatched blastocysts and isolated inner cell masses (ICM) from intermediate and late hatched blastocysts on feeder layers prepared from permanent mouse embryonic fibroblasts (STO). The cells of these porcine embryo-derived cell lines had a morphology similar to that of murine ES cells, but colony morphology was more epitheliallike. The cell lines retained a normal diploid karyotype, consistently expressed alkaline phosphatase activity, and survived cryopreservation. When subjected to in vitro differentiation, either spontaneous or induced, the embryo-derived cell lines differentiated extensively into a wide range of cell types representing the 3 embryonic germ layers. In vivo pluripotency of the cells was demonstrated by birth of a chimeric piglet, documented by pigmentation and DNA markers, and the ability to direct the development of nuclear-transfer embryos to the blastocyst stage. Such pluripotent embryo-derived cells provide a potential route for porcine genetic manipulation,
ISSN:0093-691X
1879-3231
DOI:10.1016/S0093-691X(99)00122-3