Method of engrafting cells from solid tissues

A cell and tissue technology, applied in the field of cell transplantation compositions, can solve the problems of low transplantation efficiency, weak effect, and poor cell survival

Inactive Publication Date: 2013-03-20
THE UNIV OF NORTH CAROLINA AT CHAPEL HILL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Indeed, when cells from solid organs are transplanted via vascular routes, they are less effective due to low engraftment efficiency, poor cell survival, and propensity to form life-threatening emboli

Method used

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  • Method of engrafting cells from solid tissues
  • Method of engrafting cells from solid tissues
  • Method of engrafting cells from solid tissues

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0104] According to the described protocol, mouse liver progenitor cells were isolated from recipient C57 / BL6 mice (4-5 weeks old). In order to "transplant" research, GFP (green fluorescent protein) reporter was introduced into the liver progenitor cells. Then, the cells are mixed with hyaluronic acid (HA) hydrogel, and the HA is cross-linked with poly(ethylene glycol)-diacrylate (PEG-DA) before the HA is introduced into the subject mouse. For introduction / implantation, the mice were anesthetized with ketamine (90mg / kg-120mg / kg) and xylazine (10mg / kg), and the mouse abdomen was opened. Then, cells with or without HA are slowly injected into the anterior liver lobe. The cutting site was closed and the animals were given buprenorphine at 0.1 mg / kg every 12 hours for 48 hours. After 48 hours, the animals were euthanized, and the tissues were removed, fixed, and sectioned for histology.

[0105] In order to determine the cell location in the murine animal model, the "control" hepa...

Embodiment 2

[0109] According to the described protocol, human hepatic progenitor cells were isolated from fetal liver tissue (16-20 weeks). An adenovirus vector expressing luciferase is introduced into the hepatic progenitor cells. Then, before introducing the cells into the subject mice, in the presence of the cross-linking agent poly(ethylene glycol)-diacrylate (PEG-DA), the cells were combined with thiol-modified carboxymethyl HA (CMHA-S) mixed. More specifically, the hydrogel is formed by dissolving the HA dry reagent in KM to obtain a 2.0% (weight / volume) solution, and the crosslinking agent is dissolved in KM to obtain a 4.0% weight / volume solution. Then, incubate the sample in a 37°C water bath to completely dissolve the sample. Collagen III and laminin were prepared at a concentration of 1.0 mg / ml and mixed with the crosslinker / hydrogel at a ratio of 1:4.

[0110] For introduction / implantation, the mice were anesthetized with ketamine (90-120 mg / kg) and xylazine (10 mg / kg), and th...

Embodiment 3

[0117] Human pancreatic progenitor cells were isolated from pancreatic tissue. An adenovirus vector expressing luciferase is introduced into the progenitor cell. As described in Example 2, in the presence of the crosslinking agent poly(ethylene glycol)-diacrylate (PEG-DA), the cells were combined with thiol-modified carboxymethyl HA (CMHA-S) mixing.

[0118] For introduction / implantation, the mice were anesthetized with ketamine (90mg / kg-120mg / kg) and xylazine (10mg / g), and the mouse abdomen was opened. Then, cells with or without HA are slowly injected into the pancreas. The cutting site was closed and the animals were given buprenorphine at 0.1 mg / kg every 12 hours for 48 hours. After 48 hours, the animals were euthanized, and the tissues were removed, fixed, and sectioned for histology.

[0119] In order to determine the cell location in the murine animal model, the "control" progenitor cells were infected with an adenovirus vector expressing luciferase at 50 POI at 37°C for...

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Abstract

A method of repairing diseased or dysfunctional organs or of establishing a model system of a disease state is provided. For repairing diseased organs, the method involves engraftment of cells from healthy tissue of the diseased or dysfunctional organ admixed with gel-forming biomaterials and nutrient medium, signaling molecules and extracellular matrix components that can be made insoluble rapidly upon transplantation to form a graft. In this way, the graft mimics the complexity of the native microenvironment with a minimum number of components that allow transplantation of cells to successfully engraft, expand and then rebuild part or the entirety of the diseased or dysfunctional organ. In the case of using grafting methods for establishing a disease model, diseased cells may be transplanted in the biomaterials and into experimental hosts.

Description

[0001] Inventors: Rachel Turner, David Gober, Oswaldo Rozoa, Lola M. Reid [0002] Cross reference to related patent applications [0003] This application claims the rights of US provisional patent application No. 61 / 332,441 filed on May 7, 2010, and the entire content of the US provisional patent application is incorporated herein by introduction. Technical field [0004] The present invention is generally directed to the field of tissue transplantation. More specifically, the present invention relates to compositions and methods for cell transplantation. Background technique [0005] The current method of cell transplantation therapy introduces donor cells into the recipient through the vascular pathway, which is a therapy that mimics hematopoietic therapy. However, since hematopoietic cells have been formed into a suspended state and have inherent characteristics that support their targeting to specific target tissues, it is relatively easy to perform hematopoietic cell therapy....

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61M31/00A61K35/39A61K35/407
CPCA61L2300/414A61F2/07A61K9/0024C12N2537/10A61K35/39C12N2533/80A61K9/00A61L27/54A61K47/34A61L2430/28A61L27/52C12N5/0672A61K35/407A61L27/38A61K38/00A61L27/00A61K47/36A61P1/00A61P1/16A61P43/00Y02A50/30A61K2300/00A61K35/12A61K35/42C12N5/00C12N5/06A61K31/734A61M37/00
Inventor 雷琪儿·特讷大卫·戈伯奥斯瓦杜·罗佐亚洛拉·M·里德
Owner THE UNIV OF NORTH CAROLINA AT CHAPEL HILL
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