Hostname: page-component-586b7cd67f-vdxz6 Total loading time: 0 Render date: 2024-11-22T20:35:15.010Z Has data issue: false hasContentIssue false
  • English
  • Français

Increased blastocyst formation of cloned porcine embryos produced with donor cells pre-treated with Xenopus egg extract and/or digitonin

Published online by Cambridge University Press:  08 February 2011

Ying Liu ,
Olga Østrup ,
Juan Li ,
Gábor Vajta ,
Lin Lin ,
Peter M. Kragh ,
Stig Purup ,
Poul Hyttel  and
Henrik Callesen
Ying Liu*
Affiliation:
Department of Genetics and Biotechnology, Faculty of Agricultural Sciences, Aarhus University, DK-8830 Tjele, Denmark.
Olga Østrup
Affiliation:
Department of Animal and Veterinary Basic Sciences, Faculty of Life Sciences, University of Copenhagen, Denmark.
Juan Li
Affiliation:
Department of Genetics and Biotechnology, Faculty of Health Science; Aarhus University, Denmark.
Gábor Vajta
Affiliation:
Department of Genetics and Biotechnology, Faculty of Health Science; Aarhus University, Denmark.
Lin Lin
Affiliation:
Department of Genetics and Biotechnology, Faculty of Health Science; Aarhus University, Denmark. Department of Human Genetics, Faculty of Health Science; Aarhus University, Denmark.
Peter M. Kragh
Affiliation:
Department of Genetics and Biotechnology, Faculty of Health Science; Aarhus University, Denmark.
Stig Purup
Affiliation:
Department of Animal Health and Bioscience, Faculty of Agricultural Sciences, Faculty of Health Science; Aarhus University, Denmark.
Poul Hyttel
Affiliation:
Department of Animal and Veterinary Basic Sciences, Faculty of Life Sciences, University of Copenhagen, Denmark.
Henrik Callesen
Affiliation:
Department of Genetics and Biotechnology, Faculty of Health Science; Aarhus University, Denmark.
*
All correspondence to: Ying Liu. Department of Genetics and Biotechnology, Faculty of Agricultural Sciences, Aarhus University, DK-8830 Tjele, Denmark. Tel: +45 8999 1149. Fax: +45 8999 1300. e-mail: [email protected]
Get access Rights & Permissions [Opens in a new window]

Summary

Pre-treating donor cells before somatic cell nuclear transfer (SCNT, ‘cloning’) may improve the efficiency of the technology. The aim of this study was to evaluate the early development of cloned embryos produced with porcine fibroblasts pre-treated with a permeabilizing agent and extract from Xenopus laevis eggs. In Experiment 1, fetal fibroblasts were permeabilized by digitonin, incubated in egg extract and, after re-sealing of cell membranes, cultured for 3 or 5 days before use as donor cells in handmade cloning (HMC). Controls were produced by HMC with non-treated donor cells. The blastocyst rate for reconstructed embryos increased significantly when digitonin-permeabilized, extract-treated cells were used after 5 days of culture after re-sealing. In Experiment 2, fetal and adult fibroblasts were treated with digitonin alone before re-sealing the cell membranes, then cultured for 3 or 5 days and used as donor cells in HMC. Treatment with digitonin alone increased the blastocyst rate, but only when fetal, and not adult fibroblasts, were used as donor cells, and only after 3 days of culture. In conclusion, we find a time window for increased efficiency of porcine SCNT using donor cells after pre-treatment with permeabilization/re-sealing and Xenopus egg extract. Interestingly, we observe a similar increase in cloning efficiency by permeabilization/re-sealing of donor cells without extract treatment that seems to depend on choice of donor cell type. Thus, pre-treatment of donor cells using permeabilizing treatment followed by re-sealing and in vitro culture for few days could be a simple way to improve the efficiency of porcine cloning.

Keywords

PermeabilizationPorcinePre-treatmentSomatic cell nuclear transferXenopus egg extract
Type
Research Article
Information
Zygote , Volume 20 , Issue 1 , February 2012 , pp. 61 - 66
Copyright
Copyright © Cambridge University Press 2011

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

References

Adam, S.A., Sterne-Marr, R. & Gerace, L. (1992). Nuclear protein import using digitonin-permeabilized cells. Methods Enzymol. 219, 97–110.CrossRefGoogle ScholarPubMed
Ahnert-Hilger, G., Wegenhorst, U., Stecher, B., Spicher, K., Rosenthal, W. & Gratz, M. (1992). Exocytosis from permeabilized bovine adrenal chromaffin cells is differently modulated by guanosine 5′-[γ-thio]triphosphate and guanosine 5’-[βγ-imido]triphosphate. Evidence for the involvement of various guanine nucleotide-binding proteins. Biochem. J. 284, 321–26.CrossRefGoogle Scholar
Alberio, R., Johnson, A.D., Stick, R. & Campbell, K.H. (2005). Differential nuclear remodeling of mammalian somatic cells by Xenopus laevis oocyte and egg cytoplasm. Exp. Cell. Res. 307, 131–41.CrossRefGoogle ScholarPubMed
Betthauser, J.M., Pfister-Genskow, M., Xu, H., Golueke, P.J., Lacson, J.C., Koppang, R.W., Myers, C., Liu, B., Hoeschele, I., Eilertsen, K.J. & Leno, G.H. (2006). Nucleoplasmin facilitates reprogramming and in vivo development of bovine nuclear transfer embryos. Mol. Reprod. Dev. 73, 977–86.CrossRefGoogle ScholarPubMed
Boquest, A.C., Day, B.N. & Prather, R.S. (1999). Flow cytometric cell cycle analysis of cultured porcine fetal fibroblast cells. Biol. Reprod. 60, 1013–9.CrossRefGoogle ScholarPubMed
Bru, T., Clarke, C., McGrew, M.J., Sang, H.M., Wilmut, I. & Blow, J.J. (2008). Rapid induction of pluripotency genes after exposure of human somatic cells to mouse ES cell extracts. Exp. Cell Res. 314, 2634–42.CrossRefGoogle ScholarPubMed
Chiang, C.Y. & Tang, P.C. (2009). Selection of pluripotent stem cells induced by Xenopus egg extracts. Reprod. Fertil. Dev. 21, 234.CrossRefGoogle Scholar
Du, Y., Kragh, P.M., Zhang, X., Purup, S., Yang, H., Bolund, L. & Vajta, G. (2005). High overall in vitro efficiency of porcine handmade cloning (HMC) combining partial zona digestion and oocyte trisection with sequential culture. Cloning Stem Cells 7, 199–205.CrossRefGoogle ScholarPubMed
Du, Y., Kragh, P.M., Zhang, Y., Li, J., Schmidt, M., Bøgh, I.B., Zhang, X., Purup, S., Jørgensen, A.L., Pedersen, A.M., Villemoes, K., Yang, H., Bolund, L. & Vajta, G. (2007). Piglets born from handmade cloning, an innovative cloning method without micromanipulation. Theriogenology 68, 1104–10.CrossRefGoogle ScholarPubMed
Du, Y., Lin, L., Schmidt, M., Bøgh, I.B., Kragh, P.M., Sørensen, C.B., Li, J., Purup, S., Pribenszky, C., Molnár, M., Kuwayama, M., Zhang, X., Yang, H., Bolund, L. & Vajta, G. (2008). High hydrostatic pressure treatment of porcine oocytes before handmade cloning improves developmental competence and cryosurvival. Cloning Stem Cells 10, 325–30.CrossRefGoogle ScholarPubMed
Grant, N.J., Aunis, D. & Bader, M.F. (1987). Morphology and secretory activity of digitonin- and alpha-toxin-permeabilized chromaffin cells. Neuroscience 23, 1143–55.CrossRefGoogle ScholarPubMed
Håkelien, A.M., Landsverk, H.B., Robl, J.M., Skålhegg, B.S. & Collas, P. (2002). Reprogramming fibroblasts to express T-cell functions using cell extracts. Nat. Biotechnol. 20, 460–6.CrossRefGoogle ScholarPubMed
Hansis, C., Barreto, G., Maltry, N. & Niehrs, C. (2004). Nuclear reprogramming of human somatic cells by Xenopus egg extract requires BRG1. Curr. Biol. 14, 1475–80.CrossRefGoogle ScholarPubMed
Higa, M.M., Ullman, K.S. & Prunuske, A.J. (2006). Studying nuclear disassembly in vitro using Xenopus egg extract. Methods 39, 284–90.CrossRefGoogle ScholarPubMed
Kragh, P.M., Vajta, G., Corydon, T.J., Purup, S., Bolund, L. & Callesen, H. (2004). Production of transgenic porcine blastocysts by hand-made cloning. Reprod. Fertil. Dev. 16, 315–8.CrossRefGoogle ScholarPubMed
Kragh, P.M., Du, Y., Corydon, T.J., Purup, S., Bolund, L. & Vajta, G. (2005). Efficient in vitro production of porcine blastocysts by handmade cloning with a combined electrical and chemical activation. Theriogenology 64, 1536–45.CrossRefGoogle ScholarPubMed
Lewis, M.L. & Hughes-Fulford, M. (2000). Regulation of heat shock protein message in Jurkat cells cultured under serum-starved and gravity-altered conditions. J. Cell Biochem. 77, 127–34.3.0.CO;2-0>CrossRefGoogle ScholarPubMed
Li, J., Du, Y., Zhang, Y.H., Kragh, P.M., Purup, S., Bolund, L., Yang, H., Xue, Q.Z. & Vajta, G. (2006). Chemically assisted handmade enucleation of porcine oocytes. Cloning Stem Cells 8, 241–50.CrossRefGoogle ScholarPubMed
Lin, L., Kragh, P.M., Purup, S., Kuwayama, M., Du, Y., Zhang, X., Yang, H., Bolund, L., Callesen, H. & Vajta, G. (2009). Osmotic stress induced by sodium chloride, sucrose or trehalose improves cryotolerance and developmental competence of porcine oocytes. Reprod. Fertil. Dev. 21, 338–44.CrossRefGoogle ScholarPubMed
Miyamoto, K., Furusawa, T., Ohnuki, M., Goel, S., Tokunaga, T., Minami, N., Yamada, M., Ohsumi, K. & Imai, H. (2007a). Reprogramming events of mammalian somatic cells induced by Xenopus laevis egg extracts. Mol. Reprod. Dev. 74, 1268–77.CrossRefGoogle ScholarPubMed
Miyamoto, K., Ohnuki, M., Minami, N., Yamada, M. & Imai, H. (2007b). Nuclear reprogramming of porcine cells and their use as donor cells for nuclear transfer after treatment in Xenopus egg extracts. Reprod. Fertil. Dev. 19, 150.CrossRefGoogle Scholar
Miyamoto, K., Yamashita, T., Tsukiyama, T., Kitamura, N., Minami, N., Yamada, M. & Imai, H. (2008). Reversible membrane permeabilization of mammalian cells treated with digitonin and its use for inducing nuclear reprogramming by Xenopus egg extracts. Cloning Stem Cells 10, 535–42.CrossRefGoogle ScholarPubMed
Miyamoto, K., Tsukiyama, T., Yang, Y., Li, N., Minami, N., Yamada, M. & Imai, H. (2009). Cell free extracts from mammalian oocytes partially induce nuclear reprogramming in somatic cells. Biol. Reprod. 80, 935–43.CrossRefGoogle ScholarPubMed
Naruse, K., Quan, Y.S., Kim, B.C., Choi, S.M., Park, C.S. & Jin, D.I. (2009). Streptolysin-O treatment of fetal fibroblasts improves cell fusion and in vitro development of porcine nuclear transfer embryos. J. Reprod. Dev. 55, 236–9.CrossRefGoogle ScholarPubMed
Tang, S., Wang, Y., Zhang, D., Gao, Y., Ma, Y., Yin, B., Sun, J., Liu, J. & Zhang, Y. (2009). Reprogramming donor cells with oocyte extracts improves in vitro development of nuclear transfer embryos. Anim. Reprod. Sci. 115, 1–9.CrossRefGoogle ScholarPubMed
Vajta, G., Holm, P., Greve, T. & Callesen, H. (1997). The submarine incubation system, a new tool for in vitro embryos culture. A technique report. Theriogenology 48, 1379–85.CrossRefGoogle Scholar
Vajta, G., Peura, T.T., Holm, P., Páldi, A., Greve, T., Trounson, A.O. & Callesen, H. (2000). New method for culture of zona-included or zona-free embryos: the Well of the Well (WOW) system. Mol. Reprod. Dev. 55, 256–64.3.0.CO;2-7>CrossRefGoogle ScholarPubMed
van de Ven, A.L., Adler-Storthz, K. & Richards-Kortum, R. (2009). Delivery of optical contrast agents using Triton-X100, part 1: reversible permeabilization of live cells for intracellular labeling. J. Biomed. Opt. 14, 021012.CrossRefGoogle ScholarPubMed
Walev, I., Bhakdi, S.C., Hofmann, F., Djonder, N., Valeva, A., Aktories, K. & Bhakdi, S. (2001). Delivery of proteins into living cells by reversible membrane permeabilization with streptolysin-O. Proc. Natl. Acad. Sci. USA 98, 3185–90.CrossRefGoogle ScholarPubMed
Yoshioka, K., Suzuki, C., Tanaka, A., Anas, I.M. & Iwamura, S. (2002) Birth of piglets derived from porcine zygotes cultured in a chemically defined medium. Biol. Reprod. 66, 112–9.CrossRefGoogle Scholar