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Efficient Derivation and Long Term Maintenance of Pluripotent Porcine Embryonic Stem-like Cells |
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Hye-Young Son, Jung-Eun Kim, Sang-Goo Lee, Hye-Sun Kim, Eugene Lee, Jin-Kyu Park, Hakhyun Ka, Hyun-Jong Kim, Chang-Kyu Lee* |
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Correspondence:
Chang-Kyu Lee,
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| Abstract |
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Porcine embryonic stem (ES) cells have a great potential as tools for transgenic animal production and studies of regulation of differentiation genes. Although several studies showed successful derivation of porcine ES-like cells, these cells were not maintained long-term in culture. Therefore, this study was conducted to establish porcine pluripotent ES-like cells using in vivo fertilized embryos and to maintain these cells in long term culture. Porcine ES-like cells from in vivo embryos obtained by immuno-surgery or whole explant culture were successfully cultured for over 56 passages. Morphology of porcine ES-like cells was flat-shaped with a monolayer type colony. These cells stained for alkaline phosphatase throughout the culture. Furthermore, porcine ES-like cells reacted with antibodies against Oct-4, SSEA-1, SSEA-4, Tra-1-60, and Tra-1-81, which are typical markers of undifferentiated stem cells. To characterize the ability of porcine ES-like cells to differentiate into three germ layers, embryoid body formation was induced. After plating of these cells, porcine ES-like cells were spontaneously differentiated into various cell types of all three germ layers. In addition, porcine ES-like cells were successfully derived from IVF blastocysts in media containing human recombinant basic fibroblast growth factor. |
| Keywords:
Porcine; Embryonic Stem Cells; Pluripotent; IVF; Basic Fibroblast Growth Factor |
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