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Primate embryonic stem cells

a technology of stem cells and embryos, applied in the field of stem cell cultures, can solve the problems of limited range of differentiation of human ec cells, pregnancy failure, and limited usefulness of mouse es cells as a model of human development, and achieve the effect of high nucleus/cytoplasm ratio

Inactive Publication Date: 2005-07-21
THOMSON
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Benefits of technology

[0016] It is an advantageous feature of the present invention that the primate ES cell lines continue to proliferate in an undifferentiated state after continuous culture for at least one year. In a particularly advantageous embodiment, the cells remain euploid after proliferation in an undifferentiated state.

Problems solved by technology

Although the study of mouse ES cells provides clues to understanding the differentiation of general mammalian tissues, dramatic differences in primate and mouse development of specific lineages limits the usefulness of mouse ES cells as a model of human development.
Without progesterone, pregnancy fails.
However, the range of differentiation of these human EC cells is limited compared to the range of differentiation obtained with mouse ES cells, and all EC cell lines derived to date are aneuploid (Andrews, et al., 1987, supra).

Method used

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Examples

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(1) Animals and Embryos

[0077] As described above, we have developed a technique for non-surgical, uterine-stage embryo recovery from the rhesus macaque and the common marmoset.

[0078] To supply rhesus embryos to interested investigators, The Wisconsin Regional Primate Research Center (WRPRC) provides a preimplantation embryo recovery service for the rhesus monkey, using the non-surgical flush procedure described above. During 1994, 151 uterine flushes were attempted from rhesus monkeys, yielding 80 viable embryos (0.53 embryos per flush attempt).

[0079] By synchronizing the reproductive cycles of several marmosets, significant numbers of in vivo produced, age-matched, preimplantation primate embryos were studied in controlled experiments for the first time. Using marmosets from the self-sustaining colony (250 animals) of the Wisconsin Regional Primate Research Center (WRPRC), we recovered 54 viable morulae or blastocysts, 7 unfertilized oocytes or degenerate embryos, and 5 empty z...

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Abstract

A purified preparation of primate embryonic stem cells is disclosed. This preparation is characterized by the following cell surface markers: SSEA-1 (−); SSEA-4 (+); TRA-1-60 (+); TRA-1-81 (+); and alkaline phosphatase (+). In a particularly advantageous embodiment, the cells of the preparation are human embryonic stem cells, have normal karyotypes, and continue to proliferate in an undifferentiated state after continuous culture for eleven months. The embryonic stem cell lines also retain the ability, throughout the culture, to form trophoblast and to differentiate into all tissues derived from all three embryonic germ layers (endoderm, mesoderm and ectoderm). A method for isolating a primate embryonic stem cell line is also disclosed.

Description

CROSS REFERENCES TO RELATED APPLICATIONS [0001] This application is a divisional of U.S. Ser. No. 08 / 591,246 which was filed on Jan. 18, 1996, and is a continuation-in-part of U.S. Ser. No. 08 / 376,327 which was filed on Jan. 20, 1995.STATEMENT REGARDING FEDERALLY SPONSORED RESEARCH [0002] This invention was made with United States government support awarded by NIH NCRR Grant No. RR00167. The United States government has certain rights in this invention.BACKGROUND OF THE INVENTION [0003] In general, the field of the present invention is stem cell cultures. Specifically, the field of the present invention is primate embryonic stem cell cultures. [0004] In general, stem cells are undifferentiated cells which can give rise to a succession of mature functional cells. For example, a hematopoietic stem cell may give rise to any of the different types of terminally differentiated blood cells. Embryonic stem (ES) cells are derived from the embryo and are pluripotent, thus possessing the capa...

Claims

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Application Information

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Patent Type & Authority Applications(United States)
IPC IPC(8): C12N5/073C12N5/0735
CPCC12N5/0603C12N5/0606C12N2506/02C12N5/0605C12N2502/13A61P25/16A61P37/00A61P3/10
Inventor THOMSON
Owner THOMSON
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