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Use and application of rat cardiac muscle cell cultured clear liquid

A technology of cardiomyocytes and culture supernatant, which is applied in the direction of embryonic cells, germ cells, tissue culture, etc., can solve the problems of high cost and poor effect of ES cells, and achieve the effect of good growth, low cost and simple preparation

Inactive Publication Date: 2005-01-26
THE FIRST AFFILIATED HOSPITAL ZHEJIANG UNIV COLLEGE OF MEDICINE +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The technical problem to be solved in the present invention is to provide a kind of low price, good effect, can keep the substratum that mouse embryonic stem cells do not produce differentiation and proliferate in in vitro culture, to solve the problem of using LIF in the prior art to add mouse ES cell culture medium to in vitro culture. Disadvantages of high cost and poor effect of mouse ES cells

Method used

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  • Use and application of rat cardiac muscle cell cultured clear liquid
  • Use and application of rat cardiac muscle cell cultured clear liquid
  • Use and application of rat cardiac muscle cell cultured clear liquid

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] 1.1 Prepare the following materials:

[0025] 1.1.1 The mouse embryonic stem cell line was donated by Dr. Robertson E.

[0026] 1.1.2 Rat cardiomyocyte culture supernatant prepared by oneself

[0027] 1.1.3 Glutamine (L-glutamine, Shanghai Sangong, Amresco product packaging, LotNo.1490B40).

[0028] 1.1.4 Fetal bovine serum

[0029] 1.1.5 Gibco Dulbecco`s Modified Eagle Medium (DMEM) High Glucose Medium

[0030] 1.1.6 EDTA-trypsin

[0031] 1.1.7 0.1% gelatin solution: Gelatin (gelatin, Huamei Bioengineering Company, Lot No.0205) was heated slightly with triple distilled water to prepare 0.1%, filtered through a 0.22 μm filter membrane, and stored at 4°C for later use.

[0032] 1.1.8 Reagents used for alkaline phosphatase staining: naphthol AS-MX phosphate; dimethylformamide was purchased from Shanghai Sangong; 2-amino-2-methyl-1,3 propanediol was imported from Shanghai Chemical Reagent Company Pack; fast blue RR, purchased from Fluka Company.

[0033] 1.1.9 Experi...

Embodiment 2

[0047] 2.1 Prepare the following materials:

[0048] 2.1 Prepare the following materials:

[0049] 2.1.1 The mouse embryonic stem cell line (mouse embryonic stem cell) was donated by Dr. Robertson E.

[0050] 2.1.2 Rat cardiomyocyte culture supernatant prepared by oneself

[0051] 2.1.3 Glutamine (L-glutamine, Shanghai Sangong, Amresco product packaging, LotNo.1490B40).

[0052] 2.1.4 Gibco Dulbecco`s Modified Eagle Medium (DMEM) High Glucose Medium

[0053] 2.1.5 Fetal bovine serum

[0054] 2.1.6 EDTA-trypsin

[0055] 2.1.7 0.1% gelatin solution: Gelatin (gelatin, Huamei Bioengineering Company, Lot No.0205) was heated slightly with triple distilled water to prepare 0.1%, filtered through a 0.22 μm filter membrane, and stored at 4°C for later use.

[0056] 2.1.8 Reagents used for alkaline phosphatase staining: naphthol AS-MX phosphate; dimethylformamide purchased from Shanghai Sangong; 2-amino-2-methyl-1,3 propanediol, imported from Shanghai Chemical Reagent Company Pack...

Embodiment 3

[0072] 3.1 Prepare the following materials:

[0073] 3.1.1 The mouse embryonic stem cell line was donated by Dr. Robertson E.

[0074] 3.1.2 Rat cardiomyocyte culture supernatant prepared by oneself

[0075] 3.1.3 Non-essential amino acids (non-essential amino acids solution, Gibco, Lot No.1127865)

[0076] 3.1.4 Gibco Dulbecco`s Modified Eagle Medium (DMEM) High Glucose Medium

[0077] 3.1.5 Fetal bovine serum

[0078] 3.1.6 0.1% gelatin solution: Gelatin (gelatin, Huamei Bioengineering Company, Lot No.0205) was heated slightly with triple distilled water to prepare 0.1%, filtered through a 0.22 μm membrane filter, and stored at 4°C for later use.

[0079] 3.1.7 Reagents used for alkaline phosphatase staining: naphthol AS-MX phosphate; dimethylformamide was purchased from Shanghai Sangong; 2-amino-2-methyl-1,3 propanediol was imported from Shanghai Chemical Reagent Company Pack; fast blue RR, purchased from Fluka Company.

[0080] 3.1.8 Experimental animals: SD rats were...

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Abstract

The invention relates to the usage of rat cardiac muscle cell cultured supernatant. The rat cardiac muscle cell cultured supernatant can be used for culture medium for proliferation with non-differentiation of rat embryonic stem cell and its preparation method. The rat cardiac muscle cell cultured supernatant can be used exclusively or by combination of auxiliary materials for rat embryonic stem cell special culture substrate. The preparation method for the culture substrate is also provided. To culture rat embryonic stem cell for proliferation without differentiation in vitro culture by means of the method can derive a better effect than leukemia inhibiting factor(LIF). The invention employs rat cardiac cell, has low cost and simple preparation process, being superior to LIF in promoting ES cell wall sticking, proliferating and cloning.

Description

1. Technical field [0001] The present invention relates to the use of rat cardiomyocyte culture supernatant and the culture medium for keeping mouse embryonic stem cells from differentiating and proliferating and its preparation method, especially the use of rat cardiomyocyte culture supernatant to prepare mouse embryonic stem cells without differentiation Proliferation medium and preparation method. 2. Background technology [0002] Embryonic stem cells (ES cells) are cells with pluripotent differentiation potential. They are easily differentiated in vitro, and cytokines need to be added to inhibit their differentiation. There are two types of methods for culturing ES cells in vitro: feeder-layer culture and feeder-free culture. The feeder layer culture method is an early method, mainly using mouse embryonic fibroblasts (MEF) or STO cell lines as feeder cells, which are treated to terminate their division and then prepared into a feeder monolayer, and ES cells are planted ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/00C12N5/0735
Inventor 李兰娟曹红翠
Owner THE FIRST AFFILIATED HOSPITAL ZHEJIANG UNIV COLLEGE OF MEDICINE
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