WCRB2014 POSTER PRESENTATIONS (1) (335 abstracts)
Incomplete reprogramming of bovine fetal fibroblasts by the stimulus of low pH*
Sang Jin Jin 1 , Tae Suk Kim 2 , Sang Ki Baek 1 , Song Yi Moon 1 , Yeoung-Gyu Ko 3 , Sung Woo Kim 4 , Hwan Hoo Seong 3 & Joon Hee Lee 1
1Animal Development and Biotechnology Group, Department of Animal Bioscience, College of Agriculture and Life Sciences, Gyeongsang National University, Jin ju, Republic of Korea; 2Institute of Agriculture and Life Science, College of Agriculture and Life Sciences, Gyeongsang National University, Jinju 660-701, Republic of Korea; 3Animal Genetic Resources Station, Namwon, Republic of Korea; 4National Institute of Animal Science, Rural Development Administration, kwonsun-Gu, Republic of Korea.
Stem cells can provide a useful tool for studying on mechanisms of embryonic development and production of transgenic animal. Although somatic cells are able to convert into reprogrammed cells by introduction of exogenous reprogramming factors, however, the efficiency of iPSC is still low. Murine somatic cells without transfection of exogenous transcription factors were easily able to be reprogrammed by stimulus of low pH. Here we attempted whether to convert bFFs into pluripotent cells by the stimulus of low pH. To examine the conversion of somatic cells into pluripotency, we prepared for various culture conditions (A:DMEM/F-1220%FBS, B:DMEM/F-1220%KOSR, C:DMEM/F-122%B27) supplemented with LIF, bFGF or LIFbFGF. After the cells were treated with pH 5.7 of HBSS for 25 min, they were plated in each culture medium. As results, colony formation was observed in only the cells derived from bFFs following the exposure of low pH. The colonies of bFFS stimulated bythe low pH in all culture conditions were positively reacted with AP staining. The number of positive AP colony significantly increased in B group when compared to those in A and C groups. Especially B group supplemented with LIF showed the highest AP positive staining (52.4%). Additionally we tested expression of pluripotent genes in the low pH stimulated cells using RT-PCR. Interestingly, OCT4, SOX2, KLF4 were slightly expressed but NANOG in the cells. These results showed that bFFs by stimulus of low pH were induced to convert into incomplete reprogrammed cells.
*This work was carried out with the support of NPR (2011-0013703), ‘Cooperative Research Program for Agriculture Science and Technology Development (PJ009418022014, PJ009117022014)’ RDA, Korea.
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