Register      Login
Reproduction, Fertility and Development Reproduction, Fertility and Development Society
Vertebrate reproductive science and technology
RESEARCH ARTICLE

001. ISOLATION AND CHARACTERISATION OF PORCINE ES CELLS

M. B. Nottle A B , I. M. Vassiliev A B , S. Vassilieva A B , L. F. S. Beebe A B , S. J. Harrison A B and S. M. McIlfatrick A B
+ Author Affiliations
- Author Affiliations

A Obstetrics and Gynaecology, University of Adelaide, Adelaide, SA, Australia

B Centre for Stem Cell Research, Robinson Institute, Adelaide, SA, Australia

Reproduction, Fertility and Development 21(9) 1-1 https://doi.org/10.1071/SRB09Abs001
Published: 26 August 2009

Abstract

Embryonic stem (ES) cells have the capacity for self renewal, can remain undifferentiated in long term culture and can contribute to all the cells in the body including the germ cells. ES cells have been isolated in mice and have also been described for humans. However despite considerable effort for more than two decades ES cells which can contribute to the germline are yet to be isolated for the pig or any domestic species for that matter. We have developed a new method for isolating porcine ES cells which uses whole embryos cultured in alpha MEM with 10% serum replacement plus additives under 5% O2. Unlike methods employed previously this method results in homogenous outgrowths whose cells resemble ES cells and which express Oct 4 and Nanog and SSEA-1 [1]. These cells can be passaged and cryopreserved repeatedly resulting in the establishment of cell lines at similar efficiencies to that reported previously for 129Sv mice [2]. These cells can form embryoid bodies and can be differentiated to various cell types representative of all three germ layers [3]. Following their injection into blastocysts these cells localise /become incorporated in the inner cell mass and can be used to produce chimaeras when these embryos are transferred to recipient animals [2]. To date we have produced chimaeric pigs from one male ES cell line [2]. These are currently being mated to demonstrate germline transmission. Future studies will examine the applicability of our method to other species commencing with mice and cattle before extending these to humans.

   (1) Nottle MB, Vassiliev I, Vassilieva S, Beebe LFS, Harrison, SJ, McIlfatrick SM, Ashman, RJ. (2009) Isolation of porcine embryonic stem cells using serum free conditions. Proceedings VIII International Conference on Pig Reproduction. 2009. Abstract  252-22. Banff, Canada.

   (2) Vassiliev I, Vassilieva S, Beebe LFS, Harrison SJ, McIlfatrick SM, Ashman RJ, Nottle, MB (2009) Production of chimaeric pigs using a porcine embryonic stem cell line. Proceedings 7th Annual Meeting. International Society for Stem Cell Research. 2009 Abstract 128. Barcelona, Spain.

   (3) Vassilieva S, Vassiliev I, Beebe LFS, Harrison SJ, McIlfatrick SM, Ashman RJ, Nottle MB. (2009) In vitro developmental potential of porcine embryonic stem cells. Proceedings 7th Annual Meeting. International Society for Stem Cell Research. 2009 Abstract 349.Barcelona, Spain.