Improved Neurobasal B27 culture medium, preparing method and application

A culture medium and culture solution technology, applied in biochemical equipment and methods, culture process, tissue culture, etc., can solve problems that cannot be used to study the influence of thyroid hormone on the differentiation of embryonic stem cells into nerve cells

Inactive Publication Date: 2016-05-11
THE FIRST HOSPITAL OF CHINA MEDICIAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] In the prior art, the commercially available NeurobasalB27 medium is mainly used for culturing embryonic stem cells. However, the commercially available NeurobasalB27 medium contains thyroid hormone, which cannot be used to study the influence of thyroid hormone on the differentiation of embryonic stem cells into nerve cells.

Method used

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  • Improved Neurobasal B27 culture medium, preparing method and application
  • Improved Neurobasal B27 culture medium, preparing method and application
  • Improved Neurobasal B27 culture medium, preparing method and application

Examples

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Effect test

Embodiment 1

[0030] The improved Neurobasal B27 medium of Example 1 contains the following components in every liter of Neurobasal broth: 0.1 g of biotin, 20 μg of corticosterone, 2 g of L-carnitine, 1 g of linoleic acid, 1 g of ethanolamine, 1 g of linolenic acid, D (+)-galactose 15g, progesterone 6.3μg, putrescine dihydrochloride 16.1g, retinyl acetate 0.1g, dethyroid hormone bovine serum albumin 2.5g, DL-α-tocopherol 1g, peroxide Hydrogenase 2.5g, DL-α-tocopheryl acetate 1g, reduced glutathione 1g, lipoic acid 47μg, superoxide dismutase 2.5g, transferrin 5g, human insulin 4g, sodium selenite 0.146 mg.

[0031] Each liter of modified NeurobasalB27 medium was prepared according to the following steps:

[0032] Step 1, weigh 0.1g of biotin, 20μg of corticosterone, 2g of L-carnitine, 1g of linoleic acid, 1g of ethanolamine, 1g of linolenic acid, 15g of D(+)-galactose, 6.3 µg of progesterone, 16.1 g of putrescine dihydrochloride, 0.1 g of retinyl acetate, 2.5 g of dethyroid hormone bovine ...

Embodiment 2

[0036] The improved Neurobasal B27 medium of embodiment 2 contains the following components in every liter of Neurobasal culture fluid: 0.09 g of biotin, 18 μg of corticosterone, 1 g of L-carnitine, 1.1 g of linoleic acid, 1.1 g of ethanolamine, and 1.1 g of linolenic acid g, D(+)-galactose 17g, progesterone 6.1μg, putrescine dihydrochloride 15g, retinol acetate 0.12g, dethyroid hormone bovine serum albumin 2.8g, DL-α-tocopherol 1.5g , catalase 3g, DL-α-tocopheryl acetate 1.2g, reduced glutathione 1.3g, lipoic acid 51μg, superoxide dismutase 2.8g, transferrin 5g, human insulin 4g, Sodium selenate 0.15mg.

[0037] Each liter of modified NeurobasalB27 medium was prepared according to the following steps:

[0038] Step 1, weigh 0.09g of biotin, 18μg of corticosterone, 1g of L-carnitine, 1.1g of linoleic acid, 1.1g of ethanolamine, 1.1g of linolenic acid, 17g of D(+)-semi Lactose, 6.1 μg of progesterone, 15 g of putrescine dihydrochloride, 0.12 g of retinyl acetate, 2.8 g of det...

Embodiment 3

[0041] The improved Neurobasal B27 medium of Example 3 contains the following components in every liter of Neurobasal broth: 0.11 g of biotin, 22 μg of corticosterone, 3 g of L-carnitine, 0.9 g of linoleic acid, 0.9 g of ethanolamine, and 1 g of linolenic acid , D(+)-galactose 14g, progesterone 7μg, putrescine dihydrochloride 17g, retinol acetate 0.09g, dethyroid hormone bovine serum albumin 2.2g, DL-α-tocopherol 0.9g, Catalase 2.2g, DL-α-tocopheryl acetate 1g, reduced glutathione 0.8g, lipoic acid 43μg, superoxide dismutase 2.3g, transferrin 4.2g, human insulin 3.2g, Sodium selenate 0.12mg.

[0042] Each liter of modified NeurobasalB27 medium was prepared according to the following steps:

[0043] Step 1, weigh 0-11g of biotin, 22μg of corticosterone, 3g of L-carnitine, 0-9g of linoleic acid, 0-9g of ethanolamine, 1g of linolenic acid, 14g of D(+) - Galactose, 7 μg of progesterone, 17 g of putrescine dihydrochloride, 0-09 g of retinyl acetate, 2-2 g of dethyroid hormone bov...

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Abstract

The invention discloses an improved Neurobasal B27 culture medium. One liter of a Neurobasal culture solution comprises 0.09-0.11 g of biotin, 18-22 micrograms of corticosterone, 1-3 g of L-carnitine, 0.9-1.1 g of linoleic acid, 0.9-1.1 g of ethanol amine, 0.9-1.1 g of linolenic acid, 13-17 g of D(+)-galactose, 6-7 micrograms of progesterone, 15-17 g of putrescine dihydrochloride, 0.08-0.12 g of retinyl acetate, 2.2-2.8 g of thyroid-hormone-removing bovine serum albumin, 0.8-1.5 g of DL-alpha-tocopherol, 2-3 g of catalase, 0.9-1.2 g of DL-alpha-tocopheryl acetate, 0.8-1.3 g of reduced glutathione, 42-52 micrograms of lipoic acid, 2-3 g of superoxide dismutase, 4-6 g of transferring, 3-5 g of human insulin and 0.12-0.16 mg of sodium selenite. The invention further discloses a preparing method and application of the improved Neurobasal B27 culture medium. The improved Neurobasal B27 culture medium can be used for researching the influence of thyroid hormones on the process that embryonic stem cells are differentiated into nerve cells.

Description

technical field [0001] The invention belongs to the technical field of cell culture, and in particular relates to an improved NeurobasalB27 medium, and also relates to a preparation method and application of the improved NeurobasalB27 medium. Background technique [0002] Embryonic stem cells (EmbryonicStemCell, ES cells) are derived from the inner cell mass (InnerCellMass, ICM) of the blastocyst before implantation, and have self-renewal ability and developmental pluripotency. The in vitro differentiation of embryonic stem cells provides a basis for studying the cellular and molecular mechanisms of early embryo development. a good model. [0003] Thyroid hormone is an essential hormone for fetal brain development. In iodine-deficient areas, maternal thyroid hormone deficiency during pregnancy due to iodine deficiency can lead to fetal growth retardation, mental retardation, movement disorders, and cretinism in severe cases. Therefore, it is of great significance to study t...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/079C12N5/0735
CPCC12N5/0618C12N2500/25C12N2500/30C12N2500/34C12N2500/36C12N2500/38C12N2500/46C12N2501/71C12N2501/998C12N2501/999C12N2506/02
Inventor 滕晓春滕卫平单忠艳张涵祎奚月
Owner THE FIRST HOSPITAL OF CHINA MEDICIAL UNIV
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