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Serum-free culture medium for stem cells and application of serum-free culture medium

A serum-free medium and stem cell technology, applied in the field of stem cell serum-free medium, can solve the problems of complex formula, affecting medical efficacy, limiting popularization, etc., and achieve the effect of clear chemical composition, excellent proliferation effect and low price

Active Publication Date: 2020-05-29
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the existing serum-free medium formula is complicated, and the recombinant protein is expensive, resulting in the cost of the required materials being several times higher than that of animal serum, which limits its popularity
In addition, under the existing culture conditions, mesenchymal stem cells will still experience slow proliferation and aging after several generations of culture, which greatly affects their medical efficacy.

Method used

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  • Serum-free culture medium for stem cells and application of serum-free culture medium
  • Serum-free culture medium for stem cells and application of serum-free culture medium
  • Serum-free culture medium for stem cells and application of serum-free culture medium

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0116] In this example, two sets of culture media were prepared, namely serum-free medium (SFM) and control medium (FBS). The culture medium of the control group is DMEM low-glucose medium added with 10% fetal bovine serum. The cultured cells are human adipose stem cells.

[0117] The following are detailed experimental and detection steps:

[0118] 1. The configuration of the culture medium (no need to coat when configuring)

[0119] The serum-free medium components of the present embodiment are as follows: add the following components in the DMEM low-sugar basal medium (GIBCO) of every 500mL, and make the concentration of the added components in the serum-free medium be:

[0120] Recombinant Human Serum Albumin 20mg / mL

[0121] Recombinant human PDGF-AA 20ng / mL

[0122] Recombinant human PDGF-BB 20ng / mL

[0123] Recombinant human bFGF 5ng / mL

[0124] Recombinant human TGF-β1 5ng / mL

[0125] Recombinant Human EGF 20ng / mL

[0126] Recombinant IGF 20ng / mL

[0127] Reco...

Embodiment 2

[0169] In this example, two sets of culture media were prepared, namely serum-free medium (SFM) and control medium (FBS). The culture medium of the control group is DMEM low-glucose medium added with 10% fetal bovine serum. The cultured cells are human adipose stem cells.

[0170] The following are detailed experimental and detection steps:

[0171] 1. Configuration of culture medium

[0172] Serum-free medium: add additional components in the DMEM low-sugar medium (GIBCO) of every 500mL, and make the concentration of the additional components in this serum-free medium be:

[0173] The hydrolysis rate of polyvinyl alcohol is 75-90%

[0174] PVC 10mg / mL

[0175] PDGF synthetic peptide 20ng / mL

[0176] Recombinant human bFGF 5ng / mL

[0177] Recombinant human TGF-β1 5ng / mL

[0178] Recombinant Human EGF 20ng / mL

[0179] Recombinant IGF 20ng / mL

[0180] Fibronectin FN synthetic peptide 5 μg / mL

[0181] Heparin 5μg / mL

[0182] Lipid Concentrate 0.1% (v / v)

[0183] Recom...

Embodiment 3

[0199] The cells are human umbilical cord stem cells, and the cell culture method is the same.

[0200] Serum-free medium: add additional components in the DMEM / F12 (GIBCO) of every 500mL, and make the concentration of the additional components in this serum-free medium be:

[0201] The hydrolysis rate of polyvinyl alcohol is 75-90%

[0202] PVC 10mg / mL

[0203] Recombinant human PDGF-AA 20ng / mL

[0204] Recombinant human FGF 5ng / mL

[0205] Recombinant human TGF-β1 5ng / mL

[0206] Recombinant Human EGF 20ng / mL

[0207] Recombinant human IGF 20ng / mL

[0208] Recombinant human fibronectin FN 5 μg / mL

[0209] Heparin 5μg / mL

[0210] Lipid Concentrate 0.1% (v / v)

[0211] Recombinant insulin 2 μg / mL

[0212] Transferrin 1 μg / mL

[0213] Sodium selenite 1ng / mL

[0214] Galactose 20mM

[0215] L-Glutamine 292mg / L

[0216] Putrescine 50μM

[0217] Progesterone 20nM

[0218] Hydrocortisone 100nM

[0219] Vitamin C 200μM

[0220] Vitamin A 50μM

[0221] Sodium bicarbona...

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Abstract

The invention discloses a serum-free culture medium for stem cells. The culture medium contains a basic culture medium component and serum substitutes suitable for the stem cells, wherein the serum substitutes include albumins, growth factors, adherence factors and an albumin substitute, the albumin substitute is capable of at least substituting partial albumins and is polyvinyl alcohol. By specifically utilizing the polyvinyl alcohol, the addition amount of the albumins in the culture medium is reduced; or by substituting the albumins with the polyvinyl alcohol, the cost of the culture mediumis lowered, and separation and purification of products is promoted; and in-vitro culture of the stem cells reaches a proliferation effect better that of a serum culture medium.

Description

technical field [0001] The invention belongs to the technical field of cell culture, and in particular relates to a serum-free culture medium for stem cells and an application thereof. Background technique [0002] As a kind of adult stem cells, mesenchymal stem cells are widely used in clinical research and various refractory diseases due to their advantages such as wide source, convenient material acquisition, low immunogenicity, and no ethical restrictions. A sufficient number of high-quality cells is a necessary condition to ensure therapeutic efficacy. However, the number of original mesenchymal stem cells is limited, and the isolated cells must be expanded and cultured in vitro to reach the order of magnitude for clinical use. [0003] At present, in the process of in vitro expansion and culture, the traditional mesenchymal stem cell culture medium is to add 5% to 20% animal serum to the basal medium. Serum is an indispensable component for cell growth, but as a heter...

Claims

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Application Information

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IPC IPC(8): C12N5/0775
CPCC12N5/0662C12N2500/90C12N2501/998C12N2501/10C12N2501/58C12N2500/50C12N2501/11C12N2501/115C12N2501/165C12N2501/105C12N2501/135C12N2501/15
Inventor 欧阳宏伟张琴章佳燕
Owner ZHEJIANG UNIV
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