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Cell co-culture method for improving proliferation activity of somatic cells

A technology of proliferative activity and somatic cells, applied in the field of cell co-culture to improve the proliferative activity of somatic cells, can solve the problems of tumor formation and inability to remove, and achieve the effects of simple steps, stable results and reliable principles

Inactive Publication Date: 2012-07-11
ZHONGSHAN OPHTHALMIC CENT SUN YAT SEN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, this method has the following defects: 1. The undifferentiated embryonic stem cells present in the transplanted cells will form tumors in the transplanted body
2. Stem cells are prone to differentiation in in vitro cell culture, and can be transformed into unnecessary cells of other germ layers, becoming mixed cells, which cannot be removed

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] The purpose of this embodiment is to obtain a sufficient amount of highly active corneal epithelial cells in a short time [note: the suicide genes used in this embodiment include thymidine kinase (Thymidine kinase, TK) gene, cytosine deaminase (Cytosine deaminase, CD ) gene, E.coli-gpt, cytochrome P450 (Cytochrome P450 / cytochrome P450reductase) gene, nitroreductase (Nitroreductase) gene, carboxypeptidase (Carboxypeptidase G2) gene, e.coli-DeoD gene (expressing purine nucleoside Phosphorylase), diphtheria toxin gene (DTA gene) and FAS gene and other suicide genes or one or more, stem cells are one or more of embryonic stem cells or adult stem cells, transfection methods include virus and One or more non-viral transfection methods, the time to activate the suicide gene is one or more times, and the drug to activate the suicide gene is acyclovir (ACV), ganciclovir (ganciclovir, GCv) , 5-fluorocytosine, 6-mercaptoxanthine, cyclophosphamide, 6-methylpurine-2-deoxyribonucleos...

Embodiment 2

[0036] The purpose of this embodiment is to obtain a sufficient amount of highly active human skin epidermal cells in a short time [note: the suicide genes used in this embodiment include thymidine kinase (Thymidine kinase, TK) gene, cytosine deaminase (Cytosine deaminase, CD) gene, E.coli-gpt, cytochrome P450 (Cytochrome P450 / cytochrome P450reductase) gene, nitroreductase (Nitroreductase) gene, carboxypeptidase (Carboxypeptidase G2) gene, e.coli-DeoD gene (expressing purine nucleus glycoside phosphorylase), diphtheria toxin gene (DTA gene) and FAS gene, etc. one or more of any suicide gene, stem cells are one or more of embryonic stem cells or adult stem cells, transfection methods include virus One or more of non-viral transfection methods, the time to activate the suicide gene is one or more times, and the drugs to activate the suicide gene are acyclovir (ACV), ganciclovir (Ganciclovir, GCv) ), 5-fluorocytosine, 6-mercaptoxanthine, cyclophosphamide, 6-methylpurine-2-deoxyri...

Embodiment 3

[0045] The purpose of this embodiment is to obtain a sufficient amount of highly active cat corneal endothelial cells in a short time. [Note: The suicide genes used in this embodiment include thymidine kinase (Thymidine kinase, TK) gene, cytosine deaminase (Cytosine deaminase, CD) gene, E.coli-gpt, cytochrome P450 (Cytochrome P450 / cytochrome P450reductase ) gene, Nitroreductase gene, Carboxypeptidase G2 gene, e.coli-DeoD gene (expressing purine nucleoside phosphorylase), diphtheria toxin gene (DTA gene) and FAS gene, etc. One or more kinds of suicide genes, stem cells are one or more kinds of embryonic stem cells or adult stem cells, the transfection method includes one or more kinds of viral and non-viral transfection methods, the activation of suicide genes The time is one or more times, and the drugs that activate the suicide gene are acyclovir (ACV), ganciclovir (ganciclovir, GCv), 5-fluorocytosine, 6-mercaptoxanthine, cyclophosphamide, 6 -Methylpurine-2-deoxyribonucleosi...

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Abstract

The invention discloses a cell co-culture method for improving the proliferation activity of somatic cells. The cell co-culture method comprises the following steps of: transfecting suicide genes into stem cells; co-culturing the somatic cells and the stem cells transfected with the suicide gene; and after the somatic cells are proliferated, starting the suicide genes in the stem cells to remove the stem cells. The stem cells carrying the suicide genes and the somatic cells are co-cultured, the proliferation activity of the somatic cells is improved at the presence of the stem cells, a small number of somatic cells can quickly grow in short time, and sufficient somatic cells are obtained; and when the number of the somatic cells is large enough, the suicide genes carried in the stem cells are started to induce the apoptosis of the stem cells, and the pure somatic cells with high proliferation activity are obtained, and do not have tumorigenicity after transplantation, so that the method is great breakthrough for cell engineering and tissue engineering, and is a new way for next-generation clinical application. The method has the advantages of reliable principle, simple steps and stable results, and is easily applied to experimental study and clinical treatment in the field of bioengineering.

Description

Technical field: [0001] The invention belongs to the field of cell engineering and tissue engineering, and in particular relates to a cell co-cultivation method for improving the proliferative activity of somatic cells. Background technique: [0002] In the field of cell engineering and tissue engineering, a certain number of seed cells-somatic cells are required, and somatic cells are required to have certain proliferative activity, both of which are crucial to the restoration and reconstruction of the structure and function of tissues and organs in the field of bioengineering. important influence. However, due to the limited ability of somatic cells to proliferate in vitro, a small amount of somatic cells cannot obtain a sufficient amount of somatic cells through proliferation, and after somatic cells are cultured in vitro for a long time, with the increase in the number of divisions, the proliferation ability gradually decreases, and the proliferation activity of the cell...

Claims

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Application Information

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IPC IPC(8): C12N5/07C12N5/10
Inventor 王智崇周强武征
Owner ZHONGSHAN OPHTHALMIC CENT SUN YAT SEN UNIV
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