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Culture medium for adipogenic differentiation of mesenchymal stem cells and application for culture medium

A technology of mesenchymal stem cells and adipogenic differentiation, 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, and achieve the effect of high differentiation efficiency, clear ingredients and safe use

Active Publication Date: 2020-11-24
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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  • Culture medium for adipogenic differentiation of mesenchymal stem cells and application for culture medium
  • Culture medium for adipogenic differentiation of mesenchymal stem cells and application for culture medium
  • Culture medium for adipogenic differentiation of mesenchymal stem cells and application for culture medium

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] The adipogenic differentiation medium for mesenchymal stem cells provided in this example consists of the following components: basic medium and additional components.

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

[0030]

[0031]

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

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

[0034]

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

Embodiment 2

[0042] This example provides verification of the effect of the mesenchymal stem cell adipogenic differentiation medium of Example 1. In this example, one experimental group (Example 1) and two control groups (Comparative Example 1, Comparative Example 2) are set. In the experiments of this example, human adipose-derived mesenchymal stem cells (AD-MSCs) were used, which were derived from the ATCC standard cell bank (PCS-500-011, Lot 80622175, ATCC), and these cells were used for all the following experiments.

[0043] 1. Experimental method

[0044] 1. Recovery of AD-MSCs

[0045] Take 1 AD-MSCs of P6 generation from the cell working bank, and dissolve it quickly in a 37°C water bath. The cells were slowly added to 10 mL of mesenchymal stem cell culture medium (ExCell Bio, ME000-N023), and after pipetting evenly, counted and centrifuged at 300 g for 5 minutes. Resuspend the cells with an appropriate amount of mesenchymal stem cell medium (ExCell Bio, ME000-N023), and press 80...

Embodiment 3

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

[0060] 1) culture medium

[0061]

[0062] 2) Adipogenic differentiation method

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

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Abstract

The invention relates to a culture medium for adipogenic differentiation of mesenchymal stem cells and an application thereof. The culture medium is characterized by comprising a basic culture mediumand an additive component, wherein the culture medium comprises: a variety of components like inorganic salt, amino acid and vitamins which are optimally designed, and components like recombinant human serum albumin, sodium carboxymethylcellulose, 3-isobutyl-1-methylxanthine, recombinant human insulin, dexamethasone, pioglitazone hydrochloride, palmitic acid, oleic acid, linoleic acid and cholesterol. According to the invention, through addition and proportioning of optimally-designed components, the capacity of the culture medium for inducing mesenchymal stem cells to differentiate into adipose cells is obviously improved, and the differentiation time is further shortened.

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 its application. Background technique [0002] Mesenchymal stem cells (MSCs), originating from the mesoderm, are a kind of pluripotent stem cells that mainly exist in connective tissue and organ interstitium, have strong self-renewal and multidirectional differentiation capabilities, and have the ability to differentiate in vitro It has the ability of adipocytes, bone cells, chondrocytes, nerve cells and other cells. [0003] Mesenchymal stem cells are rich in sources and widely exist in bone marrow, fat, umbilical cord and other tissues. They are convenient in source, easy to separate, culture, expand and purify, and still have stem cell characteristics after multiple passages and expansion, and have low immunogenicity. The rejection of allogeneic transplantation is relatively light, and the requirem...

Claims

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Application Information

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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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