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Aggregation of cloned embryos in empty zona pellucida improves derivation efficiency of pig ES-like cells

Published online by Cambridge University Press:  03 October 2016

Dong-Kyung Lee ,
Chi-Hun Park ,
Kwang-Hwan Choi ,
Yeon-ik Jeong ,
Kyung-Jun Uh ,
Jae Yeon Hwang ,
Sang-Goo Lee  and
Chang-Kyu Lee
Dong-Kyung Lee
Affiliation:
Department of Agricultural Biotechnology, Animal Biotechnology Major, and Research Institute of Agriculture and Life Science, Seoul National University, Seoul, Korea
Chi-Hun Park
Affiliation:
Institute of Green Bio Science and Technology, Seoul National University, Pyeong Chang, Kangwon do, 232–916, Korea
Kwang-Hwan Choi
Affiliation:
Department of Agricultural Biotechnology, Animal Biotechnology Major, and Research Institute of Agriculture and Life Science, Seoul National University, Seoul, Korea
Yeon-ik Jeong
Affiliation:
Sooam Biotech Research Foundation, Seoul 152–895, Republic of Korea
Kyung-Jun Uh
Affiliation:
Institute of Green Bio Science and Technology, Seoul National University, Pyeong Chang, Kangwon do, 232–916, Korea
Jae Yeon Hwang
Affiliation:
Department of Agricultural Biotechnology, Animal Biotechnology Major, and Research Institute of Agriculture and Life Science, Seoul National University, Seoul, Korea
Sang-Goo Lee
Affiliation:
Division of Genetics, Department of Medicine, Brigham and Women's Hospital, Harvard Medical School, Boston, MA 02115, USA
Chang-Kyu Lee*
Affiliation:
Department of Agricultural Biotechnology, Animal Biotechnology Major, and Research Institute for Agriculture and Life Science, Seoul National University, Seoul 151–921, Korea
*
All correspondence to Chang-Kyu Lee. Department of Agricultural Biotechnology, Animal Biotechnology Major, and Research Institute for Agriculture and Life Science, Seoul National University, Seoul 151–921, Korea. Tel: +1 82 2 880 4805. Fax: +1 82 2 873 4805. E-mail: leeck@snu.ac.kr
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Summary

The development of embryonic stem cells (ESCs) from large animal species has become an important model for therapeutic cloning using ESCs derived by somatic cell nuclear transfer (SCNT). However, poor embryo quality and blastocyst formation have been major limitations for derivation of cloned ESCs (ntESCs). In this study, we have tried to overcome these problems by treating these cells with histone deacetylase inhibitors (HDACi) and aggregating porcine embryos. First, cloned embryos were treated with Scriptaid to confirm the effect of HDACi on cloned embryo quality. The Scriptaid-treated blastocysts showed significantly higher total cell numbers (29.50 ± 2.10) than non-treated blastocysts (22.29 ± 1.50, P < 0.05). Next, cloned embryo quality and blastocyst formation were analyzed in aggregates. Three zona-free, reconstructed, four-cell-stage SCNT embryos were injected into the empty zona of hatched parthenogenetic (PA) blastocysts. Blastocyst formation and total cell number of cloned blastocysts increased significantly for all aggregates (76.4% and 83.18 ± 8.33) compared with non-aggregates (25.5% and 27.11 ± 1.67, P < 0.05). Finally, aggregated blastocysts were cultured on a feeder layer to examine the efficiency of porcine ES-like cell derivation. Aggregated blastocysts showed a higher primary colony formation rate than non-aggregated cloned blastocysts (17.6 ± 12.3% vs. 2.2 ± 1.35%, respectively, P < 0.05). In addition, derived ES-like cells showed typical characters of ESCs. In conclusion, the aggregation of porcine SCNT embryos at the four-cell stage could be a useful technique for improving the development rate and quality of porcine-cloned blastocysts and the derivation efficiency of porcine ntESCs.

Keywords

AggregationEmpty zonaPorcine embryoSomatic cell nuclear transfer
Type
Research Article
Information
Zygote , Volume 24 , Issue 6 , December 2016 , pp. 909 - 917
Copyright
Copyright © Cambridge University Press 2016 

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References

Biswas, D. & Hyun, S.H. (2011). Supplementation with vascular endothelial growth factor during in vitro maturation of porcine cumulus oocyte complexes and subsequent developmental competence after in vitro fertilization. Theriogenology 76, 153–60.Google Scholar
Boiani, M., Eckardt, S., Leu, N.A., Scholer, H.R. & McLaughlin, K.J. (2003). Pluripotency deficit in clones overcome by clone-clone aggregation: epigenetic complementation? The EMBO J. 22, 5304–12.Google Scholar
Choi, K.H., Park, J.K., Kim, H.S., Uh, K.J., Son, D.C. & Lee, C.K. (2013). Epigenetic changes of lentiviral transgenes in porcine stem cells derived from embryonic origin. PLoS One 8, e72184.Google Scholar
Cibelli, J.B., Stice, S.L., Golueke, P.J., Kane, J.J., Jerry, J., Blackwell, C., Ponce de Leon, F.A. & Robl, J.M. (1998). Transgenic bovine chimeric offspring produced from somatic cell-derived stem-like cells. Nat. Biotechnol. 16, 642–6.Google Scholar
Funahashi, H., Cantley, T.C. & Day, B.N. (1997). Synchronization of meiosis in porcine oocytes by exposure to dibutyryl cyclic adenosine monophosphate improves developmental competence following in vitro fertilization. Biol. Reprod. 57, 49–53.Google Scholar
Gambini, A., De Stefano, A., Bevacqua, R.J., Karlanian, F. & Salamone, D.F. (2014). The aggregation of four reconstructed zygotes is the limit to improve the developmental competence of cloned equine embryos. PLoS One 9, e110998.Google Scholar
Gambini, A., Jarazo, J., Olivera, R. & Salamone, D.F. (2012). Equine cloning: in vitro and in vivo development of aggregated embryos. Biol. Reprod. 87, 15, 11–9.Google Scholar
Gurdon, J.B. & Colman, A. (1999). The future of cloning. Nature 402, 743–6.Google Scholar
Hwang, J.Y., Mulligan, B.P., Kim, H.M., Yang, B.C. & Lee, C.K. (2013). Quantitative analysis of sperm mRNA in the pig: relationship with early embryo development and capacitation. Reprod. Fertil. Dev. 25, 807–17.Google Scholar
Koo, O.J., Park, H.J., Kwon, D.K., Kang, J.T., Jang, G. & Lee, B.C. (2009). Effect of recipient breed on delivery rate of cloned miniature pig. Zygote 17, 203–7.Google Scholar
Lee, S.G., Park, C.H., Choi, D.H., Kim, H.S., Ka, H.H. & Lee, C.K. (2007). In vitro development and cell allocation of porcine blastocysts derived by aggregation of in vitro fertilized embryos. Mol. Reprod. Dev. 74, 1436–45.Google Scholar
Misica-Turner, P.M., Oback, F.C., Eichenlaub, M., Wells, D.N. & Oback, B. (2007). Aggregating embryonic but not somatic nuclear transfer embryos increases cloning efficiency in cattle. Biol. Reprod. 76, 268–78.Google Scholar
Mombaerts, P. (2003). Therapeutic cloning in the mouse. Proc. Natl Acad. Sci. USA 100 (Suppl 1), 11924–5.Google Scholar
Munsie, M.J., Michalska, A.E., O'Brien, C.M., Trounson, A.O., Pera, M.F. & Mountford, P.S. (2000). Isolation of pluripotent embryonic stem cells from reprogrammed adult mouse somatic cell nuclei. Curr. Biol. 10, 989–92.Google Scholar
Nagy, A., Rossant, J., Nagy, R., Abramow-Newerly, W. & Roder, J.C. (1993). Derivation of completely cell culture-derived mice from early-passage embryonic stem cells. Proc. Natl Acad. Sci. USA 90, 8424–8.Google Scholar
Naruse, K., Quan, Y.S., Kim, B.C., Lee, J.H., Park, C.S. & Jin, D.I. (2007). Brief exposure to cycloheximide prior to electrical activation improves in vitro blastocyst development of porcine parthenogenetic and reconstructed embryos. Theriogenology 68, 709–16.Google Scholar
Park, C.-H., Jeong, Y.H., Jeong, Y.-I., Lee, S.-Y., Jeong, Y.-W., Shin, T., Kim, N.-H., Jeung, E.-B., Hyun, S.-H., Lee, C.-K., Lee, E. & Hwang, W.S. (2012). X-linked gene transcription patterns in female and male in vivo, in vitro and cloned porcine individual blastocysts. PLoS One 7, e51398.Google Scholar
Park, C.H., Jeong, Y.H., Lee, D.K., Hwang, J.Y., Uh, K.J., Yeom, S.C., Ahn, C. & Lee, C.K. (2014). Availability of empty zona pellucida for generating embryonic chimeras. PLoS One 10, e0123178.Google Scholar
Park, J.K., Kim, H.S., Uh, K.J., Choi, K.H., Kim, H.M., Lee, T., Yang, B.C., Kim, H.J., Ka, H.H., Kim, H. & Lee, C.K. (2013). Primed pluripotent cell lines derived from various embryonic origins and somatic cells in pig. PLoS One 8, e52481.Google Scholar
Shan, Z.Y., Wu, Y.S., Shen, X.H., Li, X., Xue, Y., Zheng, Z., Wang, Z.D., Liu, C.J., Sun, R.Z., Li, Z.Y., Shen, J.L., Liu, Z.H. & Lei, L. (2012). Aggregation of pre-implantation embryos improves establishment of parthenogenetic stem cells and expression of imprinted genes. Dev. Growth Diff. 54, 481–8.Google Scholar
Siriboon, C., Lin, Y.H., Kere, M., Chen, C.D., Chen, L.R., Chen, C.H., Tu, C.F., Lo, N.W. & Ju, J.C. (2015). Putative porcine embryonic stem cell lines derived from aggregated four-celled cloned embryos produced by oocyte bisection cloning. PLoS One 10, e0118165.Google Scholar
Siriboon, C., Tu, C.F., Kere, M., Liu, M.S., Chang, H.J., Ho, L.L., Tai, M.E., Fang, W.D., Lo, N.W., Tseng, J.K. & Ju, J.C. (2014). Production of viable cloned miniature pigs by aggregation of handmade cloned embryos at the 4-cell stage. Reprod. Fertil. Dev. 26, 395–406.Google Scholar
Sugimura, S., Narita, K., Yamashiro, H., Sugawara, A., Shoji, T., Terashita, Y., Nishimori, K., Konno, T., Yoshida, M. & Sato, E. (2009). Interspecies somatic cell nucleus transfer with porcine oocytes as recipients: A novel bioassay system for assessing the competence of canine somatic cells to develop into embryos. Theriogenology 72, 549–59.Google Scholar
Tachibana, M., Amato, P., Sparman, M., Gutierrez, N.M., Tippner-Hedges, R., Ma, H., Kang, E., Fulati, A., Lee, H.S., Sritanaudomchai, H., Masterson, K., Larson, J., Eaton, D., Sadler-Fredd, K., Battaglia, D., Lee, D., Wu, D., Jensen, J., Patton, P., Gokhale, S., Stouffer, R.L., Wolf, D. & Mitalipov, S. (2013). Human embryonic stem cells derived by somatic cell nuclear transfer. Cell 153, 1228–1238.Google Scholar
Terashita, Y., Sugimura, S., Kudo, Y., Amano, R., Hiradate, Y. & Sato, E. (2011). Improving the quality of miniature pig somatic cell nuclear transfer blastocysts: aggregation of SCNT embryos at the four-cell stage. Reprod. Domest. Anim. (Zuchthygiene) 46, 189–96.Google Scholar
Wakayama, T. (2004). On the road to therapeutic cloning. Nat. Biotechnol. 22, 399–400.CrossRefGoogle ScholarPubMed
Wen, B.Q., Li, J., Li, J.J., Tian, S.J., Sun, S.C., Qi, X., Cai, W.T. & Chang, Q.L. (2014). The histone deacetylase inhibitor Scriptaid improves in vitro developmental competence of ovine somatic cell nuclear transferred embryos. Theriogenology 81, 332–9.Google Scholar
Wu, X., Li, Y., Li, G.P., Yang, D., Yue, Y., Wang, L., Li, K., Xin, P., Bou, S. & Yu, H. (2008). Trichostatin A improved epigenetic modifications of transfected cells but did not improve subsequent cloned embryo development. Anim. Biotechnol. 19, 211–24.Google Scholar
Zhao, J., Ross, J.W., Hao, Y., Spate, L.D., Walters, E.M., Samuel, M.S., Rieke, A., Murphy, C.N. & Prather, R.S. (2009). Significant improvement in cloning efficiency of an inbred miniature pig by histone deacetylase inhibitor treatment after somatic cell nuclear transfer. Biol. Reprod. 81, 525–30.Google Scholar
Zhou, W., Xiang, T., Walker, S., Abruzzese, R.V., Hwang, E., Farrar, V., Findeisen, B., Sadeghieh, S., Arenivas, F., Chen, S.H. & Polejaeva, I. (2008). Aggregation of bovine cloned embryos at the four-cell stage stimulated gene expression and in vitro embryo development. Mol. Reprod. Dev. 75, 1281–9.Google Scholar