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Method for generating immune-compatible cells and tissues using nuclear transfer techniques

A cell and tissue technology used in the field of verifying or evaluating the immunocompatibility of these tissues to avoid transplant rejection

Inactive Publication Date: 2002-10-23
ADVANCED CELL TECH INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

This patented technology allows us to quickly find out which parts are involved with certain types of organisms called mitochruthenium (mitochloridium) genetic material. These areas may have been previously found on other individuals' chromosomes during their lifetime but they were more likely due to environmental factors like radiation exposure than usual. They could also help prevent diseases such as autoimmunity caused by bacteria or viruses.

Problems solved by technology

Technological Problem addressed in this patents relates to improving methods used during regenerative medicine procedures involving creation of various kinds of tisculites containing certain special nuclear species involved in fusion between somatic and diploepigraphy primordian neurons. Current approaches involve manipulating individual cells obtained through culture processes without specifically identifying particular ones among others, resulting in potential compounds being rejected by other normal cells. Additionally, current methodologies cannot efficiently produce isolated populations of tissile materials needed for use in future therapies targeting diseases caused by abnormalities associated with degenerating tissue functions.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0070] This experiment was designed to test the immunocompatibility of cells produced by nuclear transfer in a large preclinical animal model: bovine (Bos tauras).

[0071] Three adult Holstein castrated cattle (approximately 500-1000 lbs) approximately 8-10 months of age were purchased from Thomas Morris Company (Maryland) and shipped to South Deerfield Ranch at the University of Massachusetts (Amherst). To obtain fibroblasts for nuclear transfer, skin biopsies were obtained from each animal through an ear incision. A plasmid expressing a reporter gene encoding enhanced green fluorescent protein (eGFP) was transfected into these cells, and transfected cells were selected with neomycin. Purified cells analyzed by PCR and / or FISH were used for nuclear transfer as previously described (Nature (1998) Biotechnol. 16:642-646, incorporated herein by reference).

[0072] Embryos with more than one cell isolated from bovine blastocyst / stem cells, or blastodisc / inner cell mass or stem...

Embodiment 2

[0076] This example is designed to test teratoma formation in an immunocompromised animal model. This example relates to the method by which nuclear transfer-producing cells from patients in need of transplantation can be grown in SCID mice or other immunocompromised animals so as to produce differentiated cells for isolation and engineering of engineered tissues for transplantation .

[0077] ES cells transfected with GFP were derived from two adult Holstein castrated cattle (two different ES cell lines were obtained from each animal). ICMs were derived from day 12 blastocysts.

[0078] Cell preparation and injection procedure:

[0079] Cells are diced (cuts of no more than about 100 cells each) and filled into 1 ml syringes, not more than 200 μl, preferably 100 μl per syringe.

[0080] The ICMS was mechanically dissociated and filled into 1 ml syringes in 100-500 μl volumes.

[0081] Cells were maintained in HECM-Hepes at room temperature.

[0082] mouse#

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Embodiment 3

[0094] In order to realize the full potential of therapeutic cloning, it will be important to reconstitute more complex tissues and organs in vitro. While cloning could eliminate or greatly alleviate the most critical problem—immune compatibility—there is the hard work of assembling these cells together to generate or regenerate functional structures.

[0095] For example, myocardial infarction is one of the most common diagnoses occurring in patients seeking medical care in Western countries. Although injecting single or small groups of cardiomyocytes can help treat small focal infarcts, this approach is useful in patients with greater ischemic injuries who are at greater risk of scarring, heart rupture, and other complications. Impossible to have value. Tissue engineering offers the possibility of organizing cells into three-dimensional cardiac muscle "patches" that can be used to repair damaged parts of the heart. For heart muscle and other relatively simple tissues such ...

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Abstract

The present invention relates to methods for preparing immunocompatible tissues and cells for transplantation and tissue engineering purposes, using nuclear transfer and cloning techniques. The invention also includes methods by which the effect of expressed transgenes and other genetic manipulations on the immunocompatibility of engineered cells and tissues can be determined.

Description

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Claims

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Application Information

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Owner ADVANCED CELL TECH INC
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