Skip to main content Accessibility help
×
Home

Somatic cell nuclear transfer using transported in vitro-matured oocytes in cynomolgus monkey

  • N. Chen (a1), S-L. Liow (a1), R. Bin Abdullah (a2), WK. Khadijah Wan Embong (a2), W-Y. Yip (a1), L-G. Tan (a3), G-Q. Tong (a3) and S-C. Ng (a1) (a3)...

Summary

Somatic cell nuclear transfer (SCNT) is not successful so far in non-human primates. The objective of this study was to investigate the effects of stimulation cycles (first and repeat) on oocyte retrieval and in vitro maturation (IVM) and to evaluate the effects of stimulation cycles and donor cell type (cumulus and fetal skin fibroblasts) on efficiency of SCNT with transported IVM oocytes. In this study, 369 immature oocytes were collected laparoscopically at 24 h following human chorionic gonadotrophin (hCG) treatment from 12 cynomolgus macaque (Macaca fascicularis) in 24 stimulation cycles, and shipped in pre-equilibrated IVM medium for a 5 h journey, placed in a dry portable incubator (37 °C) without CO2 supplement. A total of 70.6% (247/350) of immature oocytes reached metaphase II (MII) stage at 36 h after hCG administration, MII spindle could be seen clearly in 80.6% (104/129) of matured IVM oocytes under polarized microscopy. A total of 50.0% (37/74) of reconstructive SCNT embryos cleaved after activation; after cleavage, 37.8% (14/37) developed to the 8-cell stage and 8.1% (3/37) developed to morula, but unfortunately none developed to the blastocyst stage. Many more oocytes could be retrieved per cycle from monkeys in the first cycle than in repeated cycles (19.1 vs. 11.7, p < 0.05). There were no significant differences in the maturation rate (70.0 vs. 71.4%, p > 0.05) and MII spindle rate under polarized microscopy (76.4 vs. 86.0%, p > 0.05) between the first and repeat cycles. There were also no significant differences in the cleavage rate, and the 4-cell, 8-cell and morula development rate of SCNT embryos between the first and repeat cycles. When fibroblast cells and cumulus cells were used as the donor cells for SCNT, first cleavage rate was not significantly different, but 4-cell (50.0 vs. 88.9%, p < 0.05) and 8-cell (0 vs. 51.9%, p < 0.01) development rate were significantly lower for the former. In conclusion, the number of stimulation cycles has a significant effect on oocyte retrieval, but has no effect on maturation and SCNT embryo development; however, different donor cell types (cumulus and fibroblast) resulted in different developmental potentials of SCNT embryos.

Copyright

Corresponding author

All correspondence to: S-C. Ng, Gleneagles Hospital, Annex Block, #01–38, 6A Napier Road, Singapore 258500, Singapore. Tel: +65 6479 7267. Fax: +65 6479 6536. e-mail: scng@embryonics.biz

References

Hide All
Arat, S., Rzucidlo, S.J., Gibbons, J., Miyoshi, K. & Stice, S.L. (2001). Production of transgenic bovine embryos by transfer of transfected granulosa cells into enucleated oocytes. Mol. Reprod. Dev. 60, 20–6.
Baguisi, A. & Overstrom, E.W. (2000). Induced enucleation in nuclear transfer procedures to produce cloned animals. Theriogenology 53, 209.
Baguisi, A., Behboodi, E., Melican, D.T., Pollock, J.S., Destrempes, M.M., Cammuso, C., Williams, J.L., Nims, S.D., Porter, C.A., Midura, P., Palacios, M.J., Ayres, S.L., Denniston, R.S., Hayes, M.L., Ziomek, C.A., Meade, H.M., Godke, R.A., Gavin, W.G., Overstrom, E.W. & Echelard, Y. (1999). Production of goats by somatic cell nuclear transfer. Nat. Biotechnol. 17, 456–61.
Bavister, B.D., Dees, C. & Schultz, R.D. (1986). Refractoriness of rhesus monkeys to repeated ovarian stimulation by exogenous gonadotropins is caused by onoprecipitating antibodies. Am. J. Reprod. Immunol. Microbiol. 11, 11–6.
Betthauser, J., Forsberg, E., Augenstein, M., Childs, L., Eilertsen, K., Enos, J., Forsythe, T., Golueke, P., Jurgella, G., Koppang, R., Lesmeister, T., Mallon, K., Mell, G., Misica, P., Pace, M., Pfister-Genskow, M., Strelchenko, N., Voelker, G., Watt, S., Thompson, S. & Bishop, M. (2000). Production of cloned pigs from in vitro systems. Nat. Biotechnol. 18, 1055–9.
Bordignon, V. & Smith, L.C. (1998). Telophase enucleation: an improved method to prepare recipient cytoplasts for use in bovine nuclear transfer. Mol. Reprod. Dev. 49, 2936.
Caligara, C., Navarro, J., Vargas, G., Simon, C., Pellicer, A. & Remohi, J. (2001). The effect of repeated controlled ovarian stimulation in donors. Hum. Reprod. 16, 2320–3.
Campbell, K.H.S. (1999). Nuclear transfer in farm animal species. Semin. Cell Dev. Biol. 10, 245–52.
Campbell, K.H.S., McWhir, J., Ritchie, W.A. & Wilmut, I. (1996). Sheep cloned by nuclear transfer from a cultured cell line. Nature 380, 64–6.
Chen, N.Q., Liow, S.L., Yip, W.Y., Tan, L.G. & Ng, S.C. (2005). Influence of cysteamine supplementation and culture in portable dry-incubator on the IVM, fertilization and subsequent development of mouse oocytes. Theriogenology 63, 2300–10.
Chen, N.Q., Liow, S.L., Abdullah, R.B., Wan Khadijah, W.E., Yip, W.Y., Tan, L.G., Tong, G.Q. & Ng, S.C. (2006). Developmental competence of transported in vitro-matured macaque oocytes. Reprod. BioMed. Online 12, 50–9.
Cibelli, J.B., Stice, S.L., Golueke, P.J., Kane, J.J., Jerry, J., Blackwell, C., León, A.P. & Robl, J.M. (1998). Cloned transgenic calves produced from non-quiescent fetal fibroblasts. Science 280, 1256–8.
Combelles, C.M. & Albertini, D.F. (2003). Assessment of oocyte quality following repeated gonadotropin stimulation in the mouse. Biol. Reprod. 68, 812–21.
Dannyes, A., De Sousa, P., King, T. & Wilmut, I. (2002). Somatic cell nuclear transfer: Recent progress and challenges. Cloning Stem Cells 4, 8190.
De Sousa, P.A., Dobrinsky, J.R., Zhu, J., Archibald, A.L., Ainslie, A., Bosma, W., Bowering, J., Bracken, J., Ferrier, P.W., Fletcher, J., Gasparrini, B., Harkness, L., Johnston, P., Ritchie, M., Ritchie, W.A., Travers, A., Albertini, D., Dinnyes, A., King, T.J. & Wilmut, I. (2002). Somatic cell nuclear transfer in pig: control of pronuclear formation and integration with improved methods for activation and maintenance of pregnancy. Biol. Reprod. 66, 642–50.
Gao, S., Chung, Y.G., William, J.W., Riley, J., Moley, K. & Latham, K.E. (2003a). Somatic cell-like features of cloned mouse embryos prepared with cultured myoblast nuclei. Biol. Reprod. 69, 4856.
Gao, S.R., McGarry, M., Ferrier, T., Pallante, B., Gasparrini, B., Fletcher, J., Harkness, L., De Sousa, P., McWhir, J. & Wilmut, I. (2003b). Effect of cell confluence on production of cloned mice using an inbred embryonic stem cell line. Biol. Reprod. 68, 595603.
Hochedlinger, K. & Jaenisch, R. (2002). Monoclonal mice generated by nuclear transfer from mature B and T donor cells. Nature 415, 1035–8.
Humpherys, D., Eggan, K., Akitsu, H., Hochedlinger, K., Rideout, W.M., Biniszkiewics, D., Yanagimachi, R. & Jaenish, R. (2001). Epignetic instability in ES cells and cloned mice. Science 293, 95–7.
Hyun, S.H., Lee, G.S., Kim, D.Y., Kim, H.S., Lee, S.H., Kim, S., Lee, E.S., Lim, J.M., Kang, S.K., Lee, B.C. & Hwang, W.S. (2003). Effect of maturation media and oocytes derived from sows or gilts on the development of cloned pig embryos. Therigenology 59, 1641–9.
Iliff, S.A., Molskness, T.A. & Stouffer, R.L. (1995). Anti-human gonadotropin antibodies generated during in vitro fertilization (IVF)-related cycles: effect on fertility of rhesus macaques. J. Med. Primatol. 24, 711.
Inoue, K., Ogonuki, N., Mochida, K., Yamamoto, Y., Takano, K., Kohto, T., Ishino, F. & Ogura, A. (2003). Effects of donor cell type and genotype on the efficiency of mouse somatic cell cloning. Biol. Reprod. 69, 1394–400.
Jain, A., Robins, J.C., Williams, D.B. & Thomas, M.A. (2005). The effect of multiple cycles in oocyte donors. Am. J. Obstet. Gynecol. 192, 1382–4.
Kasinathan, P., Knott, J.G., Wang, Z., Jerry, D.J. & Robl, J.M. (2001). Production of calves from G1 fibroblasts. Nature Biotechnol. 19, 1176–8.
Keefer, C.L., Baldassarre, H., Keyston, R., Wang, B., Bhatia, B., Bilodeau, A.S., Zhou, J.F., Leduc, M., Downey, B.R., Lazaris, A. & Karatzas, C.N. (2001). Generation of dwarf goat (Capra hircus) clones following nuclear transfer with transfected and nontransfected fetal fibroblasts and in vitro-matured oocytes. Biol. Reprod. 64, 849–56.
Kühholzer, B., Hawley, T.J., Lai, L., Kolber-Simonds, D. & Prather, R.S. (2001). Clonal lines of transgenic fibroblast cells derived from the same fetus result in different development when used for nuclear transfer in pigs. Biol. Reprod. 64, 1695–8.
Kurosaka, S., Nagao, Y., Minami, N., Yamada, M. & Imai, H. (2002). Dependence of DNA synthesis and in vitro development of bovine nuclear transfer embryos on the stage of the cell cycle of donor cells and recipient cytoplasts. Biol. Reprod. 67, 643–7.
Lanzendorf, Z.E., Zelinski-Wooten, M.B., Stouffer, R.L. & Wolf, D.P. (1990). Maturity at collection and the developmental potential of rhesus monkey oocytes. Biol. Reprod. 42, 703–11.
Li, L.Y., Connelly, M.C., Wetmore, C., Curran, T. & Morgan, J.I. (2003). Mouse embryos cloned from brain tumors. Cancer Research 63, 2733–6.
Liow, S.L., Martelli, B., Chen, N.Q., Yip, W.Y., Tan, L.G., Martelli, P., Oh, S.H., Oh, S. & Ng, S.C. (2002). Ovarian response to gonadotropin stimulation in the cynomolgus monkey, Macaca fascicularis. The Second International Symposium on Assisted Reproductive Technology (ART) for the Conservation and Genetic Management of Wildlife, 28–29 September, Omaha, Nebraska, USA.
Mitalipov, S.M., Yeoman, R.R., Nusser, K.D. & Wolf, D.P. (2002). Rhesus monkey embryos produced by nuclear transfer from embryonic blastomeres or somatic cells. Biol. Reprod. 66, 1367–73.
Miyoshi, K., Rzucidlo, S.J., Gibbons, J.R., Arat, S. & Stice, S.L. (2001). Development of porcine embryos reconstituted with somatic cells and enucleated metaphase I and II oocytes matured in a protein-free medium. BMC Dev. Biol. 1, 12.
Miyoshi, K., Rzucidlo, S.J., Pratt, S.L. & Stice, S.L. (2003). Improvements in cloning efficiencies may be possible by increasing uniformity in recipient oocytes and donor cells. Biol. Reprod. 68, 1079–86.
Ng, S.C., Martelli, P., Liow, S.L., Herbert, S. & Oh, S.H. (2002). Intracytoplasmic injection of frozen-thawed epididymal spermatozoa in a nonhuman primate model, the cynomolgus monkey (Macaca fascicularis). Theriogenology 58, 1385–97.
Ng, S.C., Chen, N.Q., Yip, W.Y., Liow, S.L., Tong, G.Q., Martelli, B., Tan, L.G. & Martelli, P. (2004). The first cell cycle after transfer of somatic cell nuclei in a non-human primate. Development 131, 2475–84.
Oback, B. & Wells, D. (2002). Donor cells for cloning: many are called but few are chosen. Cloning Stem Cells 4, 147–69.
Ono, Y., Shimozawa, N., Ito, M. & Kono, T. (2001). Cloned mice from fetal fibroblast cells arrested at metaphase by a serial nuclear transfer. Biol. Reprod. 64, 4450.
Ottobre, J.S. & Stouffer, R.L. (1985). Antibody production in rhesus monkeys following prolonged administration of human chorionic gonadotropins. Fertil. Steril. 33, 151–7.
Powell, A.M., Talbot, N.C., Wells, K.D., Kerr, D.E., Pursel, V.G. & Wall, R.J. (2004). Cell donor influences success of producing cattle by somatic cell nuclear transfer. Biol. Reprod. 71, 210–6.
Renard, J.P., Zhou, Q., LeBourhis, D., Chavatte-Palmer, P., Hue, I., Heyman, Y. & Vignon, X. (2002). Nuclear transfer technologies: between successes and doubts. Theriogenology 57, 203–22.
Schramm, R.D. & Bavister, B.D. (1999). A macaque model for studying mechanisms controlling oocyte development and maturation in human and non-human primates. Hum. Reprod. 14, 2544–55.
Schuetz, A.W., Whittingham, D.G. & Snowden, R. (1996). Alteration in the cell cycle of mouse cumulus granulusa cells during expansion and magnification in vivo and in vitro. Reprod. Fertil. Dev. 8, 935–43.
VandeVoort, C.A., Baughman, W.L. & Stouffer, R.L. (1989). Comparison of different regimens of human gonadotropins for superovulation of rhesus monkeys: ovulatory response and subsequent luteal function. J. In Vitro Fertil. Embryo Transfer 6, 8591.
VandeVoort, C.A., Leibo, S.P. & Tarantal, A.F. (2003). Improved collection and developmental competence of immature macaque oocytes. Theriogenology 59, 699707.
Vignon, X., Chesne, P., Le Bourhis, D., Flechon, J.E., Heyman, Y. & Renard, J.P. (1998). Developmental potential of bovine embryos reconstructed from enucleated matured oocytes fused with cultured somatic cells. C R Acad. Sci. III 321, 735–45.
Wakayama, T., Rodriguez, I., Perry, A.C., Yanagimachi, R. & Mombaerts, P. (1999). Mice cloned from embryonic stem cells. Proc. Natl. Acad. Sci. USA 96, 14984–9.
Wells, D.N., Misica, P.M., Day, T.A. & Tervit, H.R. (1997). Production of cloned lambs from an established embryonic cell line: a comparison between in vivo- and in vitro-matured cytoplasts. Biol. Reprod. 57, 385–93.
Wells, D.N., Pavla, P.M., Tervit, H.R. & Vivanco, W.H. (1998a). Adult somatic cell nuclear transfer is used to preserve the last surviving cow of the Enderby Island cattle breed. Reprod. Fertil. Dev. 10, 369–78.
Wells, D.N., Misica, P.M., Day, A.M., Peterson, A.J. & Tervit, H.R. (1998b). Cloning sheep from cultured embryonic cells. Reprod. Fertil. Dev. 10, 615–26.
Wells, D., Oliver, J., Miller, A., Forsyth, J., Berg, M. & Cockrem, K. (2001). Bovine cells selected in G1 of the cell cycle are totipotent following somatic cell nuclear transfer. In Proc. 32nd Ann. Conf. Soc. Reprod. Biol. Abstract No. 25.
Wells, D.N., Laible, G., Tucker, F.C., Miller, A.L., Oliver, J.E., Xiang, T., Forsyth, J.T., Berg, M.C., Cockrem, K., L'Huillier, P.J., Tervit, H.R. & Oback, B. (2003). Coordination between donor cell type and cell cycle stage improves nuclear cloning efficiency in cattle. Theriogenology 59, 4559.
Wilmut, I., Schnieke, A.E., McWhir, J., Kind, A.J. & Campbell, K.H. (1997). Viable offspring derived from fetal and adult mammalian cells. Nature 385, 810–3.
Wolf, D.P., Vandevoort, C.A., Meyer-Haas, G.R., Zelinski-Wooten, M.B., Hess, D.L., Baughman, W.L. & Stouffer, R.L. (1989). In vitro fertilization and embryo transfer in rhesus monkeys. Biol. Reprod. 41, 335–46.
Wolf, D.P., Alexander, M., Zelinski-Wooten, M. & Stouffer, R.L. (1996). Maturity and fertility of rhesus monkey oocytes collected at different intervals after an ovulatory stimulus (human chorionic gonadotrophin) in in vitro fertilized cycles. Mol. Reprod. Dev. 43, 7681.
Wolf, D.P., Meng, L., Ouhibi, O. & Zelinski-Wooten, M. (1999). Nuclear transfer in the rhesus monkey: practical and basic implications. Biol. Reprod. 60, 199204.
Wakayama, T., Rodriguez, I., Perry, A.C.F., Yanagimachi, R. & Mombaerts, P. (1999). Mice cloned from embryonic stem cells. Proc. Natl. Acad. Sci. USA 96, 14984–9.
Wakayama, T. & Yanagimachi, R. (2001a). Mouse cloning with nucleus donor cells of different age and type. Mol. Reprod. Dev. 58, 376–83.
Wakayama, T. & Yanagimachi, R. (2001b). Effect of cytokinesis inhibitors, DMSO and the timing of oocyte activation on mouse cloning using cumulus cell nuclei. Reproduction 122, 4960.
Zawada, W.M., Cibelli, J.B., Choi, P.K., Clarkson, E.D., Golueke, P.J., Witta, S.E., Bell, K.P., Kane, J., Ponce de Leon, F.A., Jerry, D.J. & Robl, J.M. (1998). Somatic cell cloned transgenic bovine neurons for transplantation in parkinsonian rats. Nat. Med. 4, 569–74.
Zelinski-Wooten, M.B., Alexander, M., Molskness, T.A., Stouffer, R.L. & Wolf, D.P. (1996). Use of recombinant human gonadotropins for repeated follicular stimulation in rhesus monkeys. In Proceedings of the XIXth conference Am. Soc. Primate. Abstract No. 133.
Zhou, Q., Jouneau, A., Brochard, V., Adenot, P. & Renard, J.P. (2001). Developmental potential of mouse embryos reconstructed from metaphase embryonic stem cell nuclei. Biol. Reprod. 65, 412–9.

Keywords

Metrics

Full text views

Total number of HTML views: 0
Total number of PDF views: 0 *
Loading metrics...

Abstract views

Total abstract views: 0 *
Loading metrics...

* Views captured on Cambridge Core between <date>. This data will be updated every 24 hours.

Usage data cannot currently be displayed