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Gynogenetic or androgenetic production of pluripotent cells and cell lines, and use thereof to produce differentiated cells and tissues

a technology of pluripotent cells and cell lines, applied in the direction of genetically modified cells, instruments, skeletal/connective tissue cells, etc., can solve the problems of never being reported in peer-reviewed journals, failure to contribute any genetic material to the resulting embryo, etc., to prevent or reduce eliminate or reduce the effect of the need for anti-rejection drugs

Inactive Publication Date: 2003-07-10
ROBL JAMES M +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

[0098] Still alternatively, the invention includes the embodiment wherein an enucleated cell, e.g., mammalian oocyte or blastomere, or other mammalian cytoplast, is transplanted with two male or female haploid nuclei, e.g., derived from oocytes or sperm, and is activated by an appropriate activation procedure to produce an embryo containing a discernible trophectoderm and inner cell mass which is incapable of giving rise to an offspring. Again, the inner cell mass or cells derived therefrom are useful for obtaining pluripotent cells which may be maintained for prolonged periods in tissue culture.
[0154] The great advantage of the subject invention is that it provides an essentially limitless supply of pluripotent, preferably pluripotent human cells that can be used to produce differentiated cells suitable for transplantation. Such cells should alleviate the significant problem associated with current transplantation methods, i.e., rejection of the transplanted tissue which may occur because of host-vs-graft or graft-vs-host rejection. Conventionally, rejection is prevented or reduced by the administration of anti-rejection drugs such as cyclosporin. However, such drugs have significant adverse side-effects, e.g., immunosuppression, carcinogenic properties, as well as being very expensive. The present invention should eliminate, or at least greatly reduce, the need for anti-rejection drugs.

Problems solved by technology

Gynogenesis includes parthenogenesis as well as activation methods wherein the spermatozoa activates the oocyte to complete meiosis, but fails to contribute any genetic material to the resulting embryo.
In fact, parthenogenetically activated oocytes may give rise to a variety of aberrations that occur during the completion of meiosis, which may result in the production of embryos of different genetic constitutions.
However, to the best of the inventors' knowledge, this work has never been reported in any peer-reviewed journal.

Method used

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  • Gynogenetic or androgenetic production of pluripotent cells and cell lines, and use thereof to produce differentiated cells and tissues
  • Gynogenetic or androgenetic production of pluripotent cells and cell lines, and use thereof to produce differentiated cells and tissues
  • Gynogenetic or androgenetic production of pluripotent cells and cell lines, and use thereof to produce differentiated cells and tissues

Examples

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example 2

Production of Parthogenic Primate Primordial Stem Cells (PPSCs)

[0184] Materials and Methods

[0185] 1--Cynomolgous Monkey (Macaca fascicularis) were superovulated using a single injection of 1000 IU of pregnant mare's serum gonadtrophin (PMSG) and 500 IU of human chorionic gonadoprophin (hCG) four days later.

[0186] 2--Ovaries were retrieved by laparotomy and oocytes dissected from the follicles and placed in maturation media 36 to 48 hrs after (hCG). Maturation media consisted of medium-199 (Gibco BRL) with Earle's balanced salt solution supplemented with 20% fetal bovine serum, 10 IU / ml of PMSG, 10 IU / ml of hCG, 0.05 mg / ml of penicillin G and 0.075 mg / ml of steptomycin sulfate (Hong, 1999).

[0187] 3--Oocyte activation. After 40 hrs in maturation, metaphase II eggs were placed in 10 micromoles of Ionomycin followed incubation in 200 mM 6-DMAP (dimethylaminopurine) for 3 to 4 hrs.

[0188] 4--Embryo culture. Commercially available embryo culture media `Cooks` was used (modified SOF). Embry...

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Abstract

Methods for obtaining pluripotent (embryonic stem) cells from parthenogenetic embryos, especially primates, are provided. These cells are useful for producing differentiated cells, tissues and organs, especially human and non-human primate cells, tissues and organs.

Description

[0001] This application is a continuation-in-part application of U.S. Ser. No. 09 / 697,297, filed Oct. 27, 2000, and further claims priority from U.S. provisional application Ser. No. 60 / 161,987, filed on Oct. 28, 1999, which are incorporated herein by reference.[0003] The present invention provides a novel method for producing pluripotent mammalian cell lines and differentiated cells, tissues and organs derived therefrom. Unlike previous reported methods for producing pluripotent cells, the subject pluripotent cells will be produced by gynogenetic or androgenetic means, i.e., methods wherein the pluripotent cells are derived from oocytes containing only DNA of male or female origin and therefore will comprise all female-derived or male-derived DNA. In a preferred embodiment, the oocytes will comprise male or female-derived DNA of primate origin, e.g. human.[0004] After ovulation, the oocytes of most mammals remain blocked at the second metaphase stage of meiosis and will eventually ...

Claims

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Application Information

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Patent Type & Authority Applications(United States)
IPC IPC(8): A61K35/12C12N5/0735C12N15/873G01N33/50
CPCA61K35/12C12N2502/13C12N15/873C12N2500/14C12N2501/999C12N2506/04C12N2510/00C12N2517/10G01N33/5008G01N33/502G01N33/5044G01N33/5073G01N33/5088G01N2333/475C12N5/0696C12N5/0606A61P43/00A61K35/545
Inventor ROBL, JAMES M.CIBELLI, JOSEBURNSIDE, AMY
Owner ROBL JAMES M
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