Abstract
To achieve human embryonic stem (ES) cell-based transplantation therapies, allogeneic transplantation models of nonhuman primates would be particularly useful. In this chapter, we describe an example of this model. We prepared cynomolgus ES cells genetically marked with the green fluorescent protein. The cells were transplanted into the allogeneic fetus because the fetus is immunologically premature and does not induce immune responses to transplanted cells. In addition, fetal tissue compartments are rapidly expanding, presumably providing space for engraftment. At 3 mo posttransplantation, a fluorescent teratoma, obviously derived from transplanted ES cells, was found in the fetus. However, transplanted cell progeny were also detected (approx 1%) in multiple fetal tissues. The cells were solitary and indistinguishable from surrounding host cells as assessed by in situ polymerase chain reaction. Transplanted cynomolgus ES cells can engraft in allogeneic fetuses. The cells will, however, form a tumor if they “leak” into an improper space, such as the thoracic cavity.
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Thomson J. A., Itskovitz-Eldor J., Shapiro S. S., et al. (1998) Embryonic stem cell lines derived from human blastocysts. Science 282, 1145–1147.
Reubinoff B. E., Pera M. F., Fong C. Y., Trounson A., and Bongso A. (2000) Embryonic stem cell lines from human blastocysts: somatic differentiation in vitro. Nat. Biotechol. 18, 399–404.
Asano T., Hanazono Y., Ueda Y., et al. (2002) Highly efficient gene transfer into primate embryonic stem cells with a simian lentivirus vector. Mol. Ther. 6, 162–168.
Lois C., Hong E. J., Pease S., Brown E. J., and Baltimore D. (2002) Germline transmission and tissue-specific expression of transgenes delivered by lentiviral vectors. Science 295, 868–872.
Pfeifer A., Ikawa M., Dayn Y., and Verma I. M. (2002) Transgenesis by lentiviral vectors: lack of gene silencing in mammalian embryonic stem cells and preimplantation embryos. Proc. Natl. Acad. Sci. USA 99, 2140–2145.
Takada T., Suzuki Y., Kondo Y., et al. (2002) Monkey embryonic stem cell lines expressing green fluorescent protein. Cell Transplant. 11, 631–635.
Harrison M. R., Slotnick R. N., Crombleholme T. M., Golbus M. S., Tarantal A. F., and Zanjani E. D. (1989) In-utero transplantation of fetal liver haemopoietic stem cells in monkeys. Lancet 2, 1425–1427.
Tarantal A. F., Goldstein O., Barley F., and Cowan M. J. (2000) Transplantation of human peripheral blood stem cells into fetal rhesus monkeys (Macaca mulatta). Transplantation 69, 1818–1823.
Suemori H., Tada T., Torii R., et al. (2001) Establishment of embryonic stem cell lines from cynomolgus monkey blastocysts produced by IVF or ICSI. Dev. Dyn. 222, 273–279.
Asano T., Ageyama N., Takeuchi K., et al. (2003) Engraftment and tumor formation after allogeneic in utero transplantation of primate embryonic stem cells. Transplantation 76, 1061–1067.
Honjo S. (1985) The Japanese Tsukuba Primate Center for Medical Science (TPC): an outline. J. Med. Primatol. 14, 75–89.
Honjo S., Cho, F., and Terao K. (1984) Establishing the cynomolgus monkey as a laboratory animal. Adv. Vet. Sci. Comp. Med. 28, 51–80.
Haase A. T., Retzel E. F., and Staskus K. A. (1990) Amplification and detection of lentiviral DNA inside cells. Proc. Natl. Acad. Sci. USA 87, 4971–4975.
Lindton B., Markling L., Ringden O., Kjaeldgaard A., Gustafson O., and Westgren M. (2000) Mixed lymphocyte culture of human fetal liver cells. Fetal Diagn. Ther. 15, 71–78.
Xu C., Inokuma M. S., Denham J., et al. (2001) Feeder-free growth of undifferentiated human embryonic stem cells. Nat. Biotechnol. 19, 971–974.
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Asano, T., Sasaki, K., Kitano, Y., Terao, K., Hanazono, Y. (2006). In Vivo Tumor Formation From Primate Embryonic Stem Cells. In: Turksen, K. (eds) Embryonic Stem Cell Protocols. Methods in Molecular Biology, vol 329. Humana Press. https://doi.org/10.1385/1-59745-037-5:459
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DOI: https://doi.org/10.1385/1-59745-037-5:459
Publisher Name: Humana Press
Print ISBN: 978-1-58829-498-2
Online ISBN: 978-1-59745-037-9
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