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Therapeutic reprogramming, hybrid stem cells and maturation

A technology of stem cells and embryonic stem cells, applied in the direction of non-embryonic pluripotent stem cells, fusion cells, animal cells, etc.

Inactive Publication Date: 2007-05-30
PRIMEGEN BIOTECH LLC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In addition, there are moral and ethical issues regarding the isolation of embryonic stem cells from human embryos

Method used

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  • Therapeutic reprogramming, hybrid stem cells and maturation
  • Therapeutic reprogramming, hybrid stem cells and maturation
  • Therapeutic reprogramming, hybrid stem cells and maturation

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0121] Maturity-pre-embryo, embryo transfer

[0122] Embryonic stem cells (ESCs) obtained from 129 / SvJ strain mice were injected into C57BL / 6J blastocysts at 3.5 days post gestation. In the blastocyst is the inner cell mass niche that contains the epiblast for the establishment of the germ layers and ultimately all cells in the embryo. ESC cells recognize this niche and respond by being properly oriented to become embryonic bodies. After a brief culture period, blastocysts are transferred back to pseudopregnant female mice and allowed to develop to term. Maturation of ESC cells directed by the inner cell mass and cellular environment into distinct stem and support cells required at specific stages in embryogenesis and organogenesis. Chimeric mice were born with varying levels of chimerism depending on the ability of the ESC cells to respond to maturation factors present during embryogenesis and organogenesis. Some of the mice had very high ESC cell contributions and some ...

Embodiment 2

[0124] Maturation of embryonic stem cells in the developing embryo

[0125] In this example, embryonic stem cells mature in the developing bone marrow niche. Blood cell development, known as hematopoiesis, proceeds through discrete stages in specific tissues in the developing embryo, before converging in the bone marrow, where it continues throughout adulthood. In the developing embryo, hematopoietic stem cell precursors first develop in the yolk sac and a region known as the aorta-gonad-mesonephros (AGM). During embryogenesis and organogenesis, hematopoietic stem cell precursors migrate to the liver, then to the spleen, and finally colonize the bone marrow before birth. In this specific example, hematopoietic, mesenchymal stem cells and multipotent adult progenitor cells (MAPCs) are generated, which can be isolated from postnatal organisms.

[0126] Embryonic stem cells (ESCs) were obtained from 129 / SvJ strain mice and transfected with a fluorescent reporter gene (ie, GFP...

Embodiment 3

[0129] Therapeutic cloning and maturation

[0130] This example describes the preparation of human primordial sex cells (donor cells) that respond to maturation signals for therapeutic cloning. In some cases, donor cells require additional steps to prepare for maturation. The methods involved in making primordial sex cells (PSCs) from other mammals, including humans, are similar to those described herein, with some possible exceptions for modifications of media or chemicals specific to that particular species.

[0131] Oogonia were harvested after ovarian stimulation and matured in vitro in G1.2 medium (Vitro Life, Goteborg, Sweden). Oogonia with first polar bodies were selected for enucleation. Ca-free buffered in HEPES 2+ Enucleation was performed in amino acid-containing CR2 medium (hCR2aa supplemented with 10% FBS and 5 μg / mL cytochalasin B (Sigma)). The oogonia are held in place with a holding pipette and a fine needle is used to create small slits in the zona pellu...

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Abstract

Therapeutically programmed cells and methods for making such cells are provided. Therapeutically programmed cells are stem cells which have been matured such that they represent either a more differentiated state or a less differentiated state after contact with stimulatory factors. The therapeutically reprogrammed cells are suitable for cellular regenerative therapy and have the potential to differentiate into more committed cell lineages. Additionally, hybrid stem cells suitable for therapeutic reprogramming and cellular regenerative therapy are provided.

Description

[0001] related application [0002] This application is U.S. Patent Application No. 10 / 346,816 filed January 16, 2003 (which claims U.S. Provisional Patent Application No. 60 / 348,521 filed January 16, 2002 and U.S. Provisional Patent Application No. Priority of U.S. Provisional Patent Application No. 60 / 367,161), a continuation-in-part of U.S. Patent Application No. 10 / 864,788 filed June 8, 2004 (which claims U.S. Provisional Patent Application No. 60 / 477,438) and claims priority to U.S. Provisional Patent Application No. 60 / 588,146, filed July 15, 2004, which is hereby incorporated by reference in its entirety. field of invention [0003] The present invention relates to the field of reprogrammed cells for therapeutic use. In particular, the present invention provides therapeutically reprogrammed cells that are not threatened by the aging process, are immunocompatible, and will function in a suitable post-natal cellular environment after implantation, to produce functional...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/16C12N5/22A61K35/12C12M3/00C12N5/00C12N5/074C12N5/076
CPCC12N5/061C12N2509/00C12N2501/235C12N2517/04A61K35/12C12N5/0611A61P11/00A61P43/00A61P9/00
Inventor 昌西·B·赛亚弗朗西斯科·J·西尔瓦
Owner PRIMEGEN BIOTECH LLC
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