430. Investigation of pluripotency in derived embryonic stem cell (ESC) lines
M. Pashai Asl A , K. Khodadadi A , M. K. Holland A , N. M. Richings A and P. J. Verma ACentre for Reproduction and Development, Monash University, Melbourne, Vic., Australia.
Reproduction, Fertility and Development 20(9) 110-110 https://doi.org/10.1071/SRB08Abs430
Published: 28 August 2008
Abstract
To produce autologous ESCs for a bovine model of cell therapy, we activated oocytes by calcium ionophore (CI) and 6 dimethylaminopurine (6 DMAP) and isolated ESCs from the resulting parthenotes. Parthenote ESC lines (pbESC) would also provide a valuable tool for epigenetic studies on ESCs. Five pbESC like-cell lines were expanded for 12 passages over 120 days and differentiated to form embryoid bodies by suspension culture. The pbESC lines demonstrated typical ESC morphology and expressed ESC markers including alkaline phosphate and stage-specific embryonic antigen, SSEA1 and SSAE4 asssessed by histochemical and immuno-fluorescence staining, respectively. In addition, gene expression of Oct4, Rex1, SSEA1 and ALP was confirmed using RT–PCR. These cells had a normal karyotype. The cells formed EBs and showed expression of the markers of three embryonic germ layers. In summary, we show that ESCs can be derived from bovine parthenogenetic blastocysts and that these cells express pluripotent markers and have ability to form EBs and differentiate into cells indicative of the three embryonic germ layers. Additional work will focus on imprinted gene expression and will provide further evidence of the parthenogenetic origin of the pbESC lines.
