A kind of mesenchymal stem cell adipogenic differentiation medium and its application

A kind of stem cell, adipogenic differentiation technology, applied in the direction of cell culture active agent, culture process, animal cells, etc., can solve the problems of long differentiation time and low differentiation efficiency, achieve high differentiation efficiency, clear components, and reduce the need for quality control Effect

Active Publication Date: 2022-07-19
苏州依科赛生物科技股份有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] The first object of the present invention is to provide a mesenchymal stem cell adipogenic differentiation medium, which can induce mesenchymal stem cells in a shorter time, in view of the shortcomings of the existing adipogenic differentiation medium, such as low differentiation efficiency and long differentiation time. differentiation into adipocytes

Method used

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  • A kind of mesenchymal stem cell adipogenic differentiation medium and its application
  • A kind of mesenchymal stem cell adipogenic differentiation medium and its application
  • A kind of mesenchymal stem cell adipogenic differentiation medium and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] The mesenchymal stem cell adipogenic differentiation medium provided in this example consists of the following components: basal medium and supplementary components.

[0029] (1) The basal medium is marked as Basal, and is prepared according to the following scheme:

[0030]

[0031]

[0032] The above-mentioned various components can be purchased from Sigma, Aladdin and other reagent companies. Each component is dissolved in 1 L of water for injection, and filtered with a 0.22-micron membrane to obtain the basic medium Basal.

[0033] (2) The added components are recorded as SUP1 components, and the composition of SUP1 components is as follows:

[0034]

[0035] In this example, 3-isobutyl-1-methylxanthine, dexamethasone, and pioglitazone hydrochloride are soluble in DMSO, while palmitic acid, oleic acid, linoleic acid, and cholesterol are soluble in absolute ethanol , recombinant human serum albumin, sodium carboxymethyl cellulose can be dissolved in water f...

Embodiment 2

[0042] This example provides verification of the effect of the mesenchymal stem cell adipogenic differentiation medium of Example 1. This example sets up one experimental group (Example 1) and two control groups (Comparative Example 1, Comparative Example 2). Human adipose-derived mesenchymal stem cells (AD-MSCs) were used in the experiments in this example, and the cells were derived from the ATCC standard cell bank (PCS-500-011, Lot 80622175, ATCC), and the cells were used in all experiments described below.

[0043] 1. Experimental method

[0044] 1. Recovery of AD-MSCs

[0045] Take 1 AD-MSCs at passage P6 from the cell working bank and dissolve them rapidly in a 37°C water bath. The cells were slowly added to 10 mL of mesenchymal stem cell medium (ExCell Bio, ME000-N023), pipetted evenly, counted, and centrifuged at 300 g for 5 minutes. Resuspend cells with an appropriate amount of mesenchymal stem cell medium (ExCell Bio, ME000-N023), and press 8000 / cm 2 The cells wer...

Embodiment 3

[0059] This example provides a test of the effect of different ratios. The basal medium (Basal) and the supplementary components (SUP1) are prepared according to the scheme of Example 1 of this example, and the basal medium and the supplementary components are prepared according to different ratios of mesenchymal stem cells. Adipogenic differentiation medium was prepared, and the effect of adipogenic induction and differentiation of each proportioning medium was tested. Oil red O staining was performed 12 days after induction of differentiation. The human adipose-derived mesenchymal stem cells and the culture method used in this example are the same as those in Example 2.

[0060] 1) Culture medium

[0061]

[0062] 2) Adipogenic induction and differentiation method

[0063] Passaging cells 20000 / cm 2 The cells were re-seeded into a 12-well plate, and the cells were cultured for 24 hours to induce differentiation. The medium described in Example 1, Comparative Example 1,...

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PUM

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Abstract

The present invention relates to a mesenchymal stem cell adipogenic differentiation medium and application thereof, which is characterized in that it comprises a basal medium and additional components, and the medium comprises: optimized design of various inorganic salts, amino acids, vitamins and other components, And recombinant human serum albumin, sodium carboxymethylcellulose, 3-isobutyl-1-methylxanthine, recombinant human insulin, dexamethasone, pioglitazone hydrochloride, palmitic acid, oleic acid, subcutaneous Oleic acid, cholesterol and other ingredients. The invention significantly improves the ability of the medium to induce the differentiation of the mesenchymal stem cells to the adipocytes by optimizing the design of the addition and ratio of the components, and further shortens the differentiation time.

Description

technical field [0001] The invention belongs to the technical field of cell culture, and in particular relates to a mesenchymal stem cell adipogenic differentiation medium and use thereof. Background technique [0002] Mesenchymal stem cells (MSCs), originating from the mesoderm, are a kind of pluripotent stem cells that mainly exist in connective tissues and organ interstitium. It is the ability of various cells such as fat cells, bone cells, cartilage cells, and nerve cells. [0003] Mesenchymal stem cells are rich in sources and widely exist in bone marrow, fat, umbilical cord and other tissues. They have convenient sources and are easy to separate, culture, expand and purify. After multiple passages and expansions, they still have stem cell characteristics and have low immunogenicity. Allograft rejection is relatively mild, and the matching requirements are not strict; it has an immunoregulatory function, and inhibits the proliferation and immune response of T cells thr...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/077
CPCC12N5/0653C12N2500/32C12N2500/38C12N2500/14C12N2500/20C12N2500/24C12N2500/40C12N2501/90C12N2501/998C12N2501/33C12N2501/39C12N2500/36C12N2501/385C12N2500/16C12N2500/12C12N2500/22C12N2500/34C12N2500/30C12N2500/46C12N2506/1384C12N2506/1353C12N2506/1369C12N2506/1392
Inventor 于宝利李其雷陈旭陈刚杨建国孙芳
Owner 苏州依科赛生物科技股份有限公司
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