Hostname: page-component-76fb5796d-45l2p Total loading time: 0 Render date: 2024-04-26T06:13:52.173Z Has data issue: false hasContentIssue false
  • English
  • Français

Isolation and initial culture of porcine inner cell masses derived from in vitro-produced blastocysts

Published online by Cambridge University Press:  01 February 2007

H-S. Kim ,
H-Y. Son ,
S. Kim ,
G-S. Lee ,
C-H. Park ,
S-K. Kang ,
B-C. Lee ,
W-S. Hwang  and
C-K. Lee
H-S. Kim
Affiliation:
Department of Theriogenology and Biotechnology, College of Veterinary Medicine, Seoul, Korea.
H-Y. Son
Affiliation:
Department of Food and Animal Biotechnology, Seoul National University, Seoul 151–742, Korea.
S. Kim
Affiliation:
Department of Theriogenology and Biotechnology, College of Veterinary Medicine, Seoul, Korea.
G-S. Lee
Affiliation:
Department of Theriogenology and Biotechnology, College of Veterinary Medicine, Seoul, Korea.
C-H. Park
Affiliation:
Department of Food and Animal Biotechnology, Seoul National University, Seoul 151–742, Korea.
S-K. Kang
Affiliation:
Department of Theriogenology and Biotechnology, College of Veterinary Medicine, Seoul, Korea.
B-C. Lee
Affiliation:
Department of Theriogenology and Biotechnology, College of Veterinary Medicine, Seoul, Korea.
W-S. Hwang
Affiliation:
Department of Theriogenology and Biotechnology, College of Veterinary Medicine, Seoul, Korea.
C-K. Lee*
Affiliation:
Department of Food and Animal Biotechnology, Seoul National University, Seoul 151–742, Korea.
*
All correspondence to: C-K. Lee, Department of Food and Animal Biotechnology, Seoul National University, Seoul 151–742, Korea. Tel: +82 2 880 4805. Fax: +82 2 873 4805. e-mail: leeck@snu.ac.kr
Get access Rights & Permissions [Opens in a new window]

Summary

The present study was conducted to isolate and culture inner cell mass (ICM) primarily derived from in vitro-produced blastocysts and to develop the culture conditions for the ICM cells. In Experiment 1, immunosurgically isolated ICMs of blastocysts derived from in vitro fertilization (IVF), somatic cell nuclear transfer (SCNT) or parthenogenetic activation (PA) were seeded onto STO cells. Primary colonies from each isolated ICM were formed with a ratio of 28.9, 30.0 and 4.9%, respectively. In Experiment 2, blastocysts collected from IVF were directly seeded onto a feeder layer with or without zona pellucida (ZP), or were subjected to ICM isolation by immunosurgery. Primary colonies were formed in 36.8% of isolated ICMs and 19.4% in intact blastocysts without ZP. In Experiment 3, ICMs from IVF blastocysts were seeded onto STO cells, mouse embryonic fibroblast (MEF) or porcine uterine epithelial cells (PUEC). On STO and MEF cells, 34.5 and 22.2% of primary colonies were formed, respectively. However, no primary colony was formed on the PUEC or in feeder-free condition. In Experiment 4, ICMs from IVF blastocysts were cultured in DMEM + Ham's F10 (D/H medium), DMEM + NCSU-23 (D/N medium) or DMEM alone. When D/H medium or D/N medium was used, 21.7 or 44.4% of primary colony were formed, respectively, while no primary colony was formed in DMEM alone. These cells showed alkaline phosphatase activity and could be maintained for up to five passages. In suspension culture, cells formed embryoid bodies. These results demonstrate that porcine ICM could be isolated and cultured primarily from in vitro-produced blastocysts with a suitable culture system.

Keywords

BlastocystsIn vitro fertilizationInner cell massPorcineSomatic cell nuclear transfer
Type
Research Article
Information
Zygote , Volume 15 , Issue 1 , February 2007 , pp. 55 - 63
Copyright
Copyright © Cambridge University Press 2007

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

Abeydeera, L.R. & Day, B.N. (1997). Fertilization and subsequent development in vitro of pig oocytes inseminated in a modified Tris-buffered medium with frozen-thawed ejaculated spermatozoa. Biol. Reprod. 57, 729–34.CrossRefGoogle Scholar
Amit, M., Shariki, C., Margulets, V. & Itskovitz-Eldor, J. (2004). Feeder and serum-free culture of human embryonic stem cells. Biol. Reprod. 70, 837–45.CrossRefGoogle ScholarPubMed
Anderson, G.B., Behboodi, E. & Pacheco, T.V. (1992). Culture of inner cell masses from in vitro derived bovine blastocysts. Theriogenology 37, 187. Abstract.CrossRefGoogle Scholar
Anderson, G.B., Choi, S.J. & BonDurant, R.H. (1994). Survival of porcine inner cell masses in culture and after injection into blastocysts. Theriogenology 42, 204–12.CrossRefGoogle ScholarPubMed
Bongso, A., Fong, C.Y., Ng, S.C. & Ratnam, S. (1994). Isolation and culture of inner cell mass cells from human blastocysts. Hum. Reprod. 9, 2110–17.CrossRefGoogle ScholarPubMed
Bradley, A., Evans, M., Kaufman, M.H. & Robertson, E. (1984). Formation of germ-line chimaeras from embryo-derived teratocarcinoma cell lines. Nature 309, 255–6.CrossRefGoogle ScholarPubMed
Chen, L.R., Shiue, Y.L., Bertolini, L., Medrano, J.F., BonDurant, R.H. & Anderson, G.B. (1999). Establishment of pluripotent cell lines from porcine preimplantation embryos. Theriogenology 52, 195–212.CrossRefGoogle ScholarPubMed
Davis, D.L. (1985). Culture and storage of pig embryos. J. Reprod. Fertil. 33, 115–24.Google Scholar
De la Fuente, R. & King, W.A. (1997). Use of a chemically defined system for the direct comparison of inner cell mass and trophectoderm distribution in murine, porcine and bovine embryos. Zygote 5, 309–20.CrossRefGoogle ScholarPubMed
Doetshman, T., Williams, P. & Maeda, N. (1988). Establishment of hamster blastocyst-derived embryonic stem (ES) cells. Dev. Biol. 127, 224–7.CrossRefGoogle Scholar
Evans, M.J. & Kaufman, M.H. (1981). Establishment in culture of pluripotential cells from mouse embryos. Nature 292, 154–6.CrossRefGoogle ScholarPubMed
Evans, M.J., Notarianni, E. & Laurie, S. (1990). Derivation and preliminary characterization of pluripotent cell lines from porcine and bovine blastocysts. Theriogenology 33, 125–8.CrossRefGoogle Scholar
Fleming, T.P., Warren, P.D., Chisholm, J.C. & Johnson, M.H. (1984). Trophectodermal processes regulate the expression of totipotency within the inner cell mass of the mouse expanding blastocyst. J. Embryol. Exp. Morphol. 84, 63–90.Google ScholarPubMed
Gallagher, E.J. & McWhir, J. (2004). The derivation of murine embryonic stem and embryonic germ cells by selective ablation. Methods Mol. Biol. 254, 99–112.Google ScholarPubMed
Graves, K.H. & Moreadith, R.W. (1993). Derivation and characterization of putative pluripotential ES cell lines from preimplantation rabbit embryos. Mol. Reprod. Dev. 36, 424–33.CrossRefGoogle ScholarPubMed
Golovan, S.P., Meidinger, R.G., Ajakaiye, A., Cottrill, M., Wiederkehr, MZ., Barney, D., Plante, C., Pollard, J., Fan, M.Z., Hayes, M.A., Laursen, J., Hjorth, J.P., Hacker, R.R., Phillips, J.P. & Forsberg, C.W. (2001). Pigs expressing salivary phytase produce low phosphorus manure. Nat. Biotech. 19, 741–5.Google Scholar
Hyun, S.H., Lee, B.C., Lee, G.S., Lee, E.S., Lim, J.M., Kang, S.K. & Hwang, W.S. (2003). Recruit of porcine oocytes excluded from nuclear transfer program for the production of embryos following parthenogenetic activation. J. Vet. Med. Sci. 65, 51–6.CrossRefGoogle ScholarPubMed
Iannaccone, P.M., Taborn, G.U., Garton, R.L., Caplice, M.D. & Brenin, D.R. (1994). Pluripotent embryonic stem cells from the rat are capable of producing chimeras. Dev. Biol. 163, 288–92.CrossRefGoogle ScholarPubMed
Kim, H.S., Lee, G.S., Hyun, S.H., Lee, S.H., Nam, D.H., Jeong, Y.W., Kim, S., Kang, S.K., Lee, B.C. & Hwang, W.S. (2004). Improved in vitro development of porcine embryos with different energy substrates and serum. Theriogenology 61, 1381–93.CrossRefGoogle ScholarPubMed
Lai, L.X., Kolber-Simonds, D., Park, K.W., Cheong, H.T., Greenstein, J.L., Im, G.S., Samuel, M., Bonk, A., Rieke, A., Day, B.N., Murphy, C.N., Carter, D.B., Hawley, R.J. & Prather, R.S. (2002). Production of alpha-1,3-galactosyltransferase knockout pigs by nuclear transfer cloning. Science 295, 1089–92.CrossRefGoogle ScholarPubMed
Lee, C.K. & Piedrahita, J.A. (2000). Effect of growth factors and feeder cells on porcine primordial germ cells in vitro. Cloning 2, 197–205.CrossRefGoogle ScholarPubMed
Lee, G.S., Hyun, S.H., Kim, H.S., Kim, D.Y., Lee, S.H., Lim, J.M., Lee, E.S., Kang, S.K., Lee, B.C. & Hwang, W.S. (2003a). Improvement of a porcine somatic cell nuclear transfer technique by optimizing donor cell and recipient oocyte preparations. Theriogenology 59, 1949–57.CrossRefGoogle ScholarPubMed
Lee, G.S., Kim, H.S., Hyun, S.H., Kim, D.Y., Lee, S.H., Nam, D.H., Jeong, Y.W., Kim, S., Kang, S.K., Lee, B.C. & Hwang, W.S. (2003b). Improved developmental competence of cloned porcine embryos with different energy supplements and chemical activation. Mol. Reprod. Dev. 66, 17–23.CrossRefGoogle ScholarPubMed
Li, M., Zhang, D., Hou, Y., Jiao, L., Zheng, X. & Wang, W.H. (2003). Isolation and culture of embryonic stem cells from porcine blastocysts. Mol. Reprod. Dev. 65, 429–34.Google Scholar
Li, M., Li, Y.H., Hou, Y., Sun, X.F., Sun, Q. & Wang, W.H. (2004). Isolation and culture of pluripotent cells from in vitro produced porcine embryos. Zygote 12, 43–8.CrossRefGoogle ScholarPubMed
Miyoshi, K., Taguchi, Y., Sendai, Y., Hoshi, H. & Sato, E. (2000). Establishment of a porcine cell line from in vitro-produced blastocysts and transfer of the cells into enucleated oocytes. Biol. Reprod. 62, 1640–6.CrossRefGoogle ScholarPubMed
Moore, K. & Piedrahita, J.A. (1996). Effects of heterologous hematopoietic cytokines on in vitro differentiation of cultured porcine inner cell masses. Mol. Reprod. Dev. 45, 139–44.3.0.CO;2-U>CrossRefGoogle ScholarPubMed
Moore, K. & Piedrahita, J.A. (1997). The effects of human leukemia inhibitory factor (hLIF) and culture medium on in vitro differentiation of cultured porcine inner cell mass (pICM). In Vitro Cell Dev. Biol. Anim. 33, 62–71.CrossRefGoogle ScholarPubMed
Mueller, S. (1999). Chimeric pigs following blastocyst injection of transgenic porcine primordial germ cells. Mol. Reprod. Dev. 54, 244–54.3.0.CO;2-5>CrossRefGoogle ScholarPubMed
Nagashima, H., Fujimura, T., Takahagib, Y., Kuromea, M., Wakoa, N., Ochiaia, T., Esakia, R., Kanoc, K., Saitod, S., Okabee, M. & Murakamib, H. (2003). Development of efficient strategies for the production of genetically modified pigs. Theriogenology 59, 95–106. Review.CrossRefGoogle ScholarPubMed
Nagashima, H., Giannakis, C., Ashman, R.J. & Nottle, M.B. (2004). Sex differentiation and germ cell production in chimeric pigs produced by inner cell mass injection into blastocysts. Biol. Reprod. 70, 702–7.CrossRefGoogle ScholarPubMed
Nichols, J., Evans, E.P. & Smith, A.G. (1990). Establishment of germ-line-competent embryonic stem (ES) cells using differentiation inhibiting activity. Development 110, 1341–8.CrossRefGoogle Scholar
Noble, M.S., Rodriguez-Zas, S., Cook, J.B., Bleck, G.T., Hurley, W.L. & Wheeler, M.B. (2002). Lactational performance of first-parity transgenic gilts expressing bovine alpha-lactalbumin in their milk. J. Anim. Sci. 80, 1090–6.CrossRefGoogle ScholarPubMed
Park, K.W., Cheong, H.T., Lai, L.X., Im, G.S., Kuhholzer, B., Bonk, A., Samuel, M., Rieke, A., Day, B.N., Murphy, C.N., Carter, D.B. & Prather, R.S. (2001). Production of nuclear transfer-derived swine that express the enhanced green fluorescent protein. Anim. Biotechnol. 12, 173–81.CrossRefGoogle ScholarPubMed
Piedrahita, J.A. (1998). Generation of transgenic porcine chimeras using primordial germ cell-derived colonies. Biol. Reprod. 58, 1321–9.CrossRefGoogle ScholarPubMed
Rosler, E.S., Fisk, G.J., Ares, X., Irving, J., Miura, T., Rao, M.S. & Carpenter, M.K. (2004). Long-term culture of human embryonic stem cells in feeder-free conditions. Dev. Dyn. 229, 259–74.Google Scholar
Silcox, R.W. & Johnson, B.H. (1988). Developmental potential of day 13 porcine embryonic disc under in vitro culture conditions. In Vitro Cell Dev. Biol. 24, 1165–72.Google Scholar
Smith, A.G., Heath, J.K., Donaldson, D.D., Wong, G.G., Moreau, J., Stahl, M. & Rogers, D. (1988). Inhibition of pluripotential embryonic stem cell differentiation by purified polypeptides. Nature 336, 688–90.Google Scholar
Soom, A.V., Boerjan, M.L., Bols, P.E.J., Vanroose, G., Lein, A., Coryn, M. & De Kruif, A. (1997). Timing of compaction and inner cell allocation in bovine embryos produced in vivo after superovulation. Biol. Reprod. 57, 1041–9.Google Scholar
Strelchenko, N. (1996). Bovine pluripotent stem cells. Theriogenology 45, 131–40.CrossRefGoogle Scholar
Strojek, R.M., Reed, M.A., Hoover, J.L. & Wagner, T.E. (1990). A method for cultivating morphologically undifferentiated embryonic stem cells from porcine blastocysts. Theriogenology 33, 901–13.CrossRefGoogle ScholarPubMed
Sukoyan, M.A., Golubitsa, A.N., Zhelezova, A.I., Shilov, A.G., Vatolin, S.Y., Maximovsky, L.P., Andreeva, L.E., McWhir, J., Pack, S.D., Bayborodin, S.I., Kerkis, A.Y., Kizilova, H.I. & Serov, O.L. (1992). Isolation and cultivation of blastocyst-derived stem cell lines from American mink (Mustela vison). Mol. Reprod. Dev. 33, 418–31.CrossRefGoogle ScholarPubMed
Talbot, N.C. & Garrett, W.M. (2001). Ultrastructure of the embryonic stem cells of the 8-day pig blastocyst before and after in vitro manipulation: development of junctional apparatus and the lethal effects of PBS mediated cell–cell dissociation. Anat. Rec. 264, 101–13.CrossRefGoogle ScholarPubMed
Thomson, J.A., Itskovitz-Eldor, J., Shapiro, S.S., Waknitz, M.A., Swiergiel, J.J., Marshall, V.S. & Jones, J.M. (1998). Embryonic stem cell lines derived from human blastocysts. Science 282, 1145–7.CrossRefGoogle ScholarPubMed
Thomson, J.A., Kalishman, J., Golos, T.G., Durning, M., Harris, C.P., Becker, R.A. & Hearn, J.P. (1995). Isolation of a primate embryonic stem cell line. Proc. Natl. Acad. Sci. USA 92, 7844–8.Google Scholar
Van De Velde, A., Liu, L., Bols, P.E., Ysebaert, M.T. & Yang, X. (1999). Cell allocation and chromosomal complement of parthenogenetic and IVF bovine embryos. Mol. Reprod. 54, 57–62.Google Scholar
Wheeler, M.B. (1994). Development and validation of swine embryonic stem cells: a review. Reprod. Fertil. Dev. 6, 563–8.Google Scholar
Wheeler, M.B. & Walters, E.M. (2001). Transgenic technology and applications in swine. Theriogenology 56, 1345–69.CrossRefGoogle Scholar
Wianny, F., Perreau, C. & Hochereau de Reviers, M.T. (1997). Proliferation and differentiation of porcine inner cell mass and epiblast in vitro. Biol. Reprod. 57, 756–64.Google Scholar
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, 112–19.CrossRefGoogle Scholar