Serum-free culture medium, preparation method thereof and culture method of mesenchymal stem cells

A serum-free medium and stem cell technology, which is applied in the field of serum-free medium and its preparation, and the culture of mesenchymal stem cells, can solve the problems of stem cell proliferation and descending ability, and achieve reduced culture cost and good biocompatibility. The effect of stability and performance

Active Publication Date: 2020-03-27
GUANGZHOU SALIAI STEMCELL SCI & TECH CO LTD
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

This patented technology describes various methods for growing or culturing mesenchyma tissue with high efficiency compared to traditional techniques like blood banking. It involves adding certain molecules called polysaccharides into the solution containing other materials known as carriers. These carrying groups enhances the stability of the microorganisms during cultivating them while maintain their functions when they come off the surface after being applied on biological surfaces. Additionally, there may exist specific types of plasma derivatives made up mostly of hydroxypropylmaleaceuclate (HPM) monomers, specifically HPM).

Problems solved by technology

This patents discusses three technical problem addressed in this patented text: 1) The challenge faced by these inventors lies in finding a commercially viable solution without relying solely on animals because many species lack natural sources like mammals while others require expensive laboratory supplies. 2) Existing methods involve either isolating pure populations of primary cells or manipulating them outside of living environments, making up even greater challenges than desired. 3) Limited availabilities of certain supplementary agents limit the effectiveness of existing commercial solutions.

Method used

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  • Serum-free culture medium, preparation method thereof and culture method of mesenchymal stem cells
  • Serum-free culture medium, preparation method thereof and culture method of mesenchymal stem cells
  • Serum-free culture medium, preparation method thereof and culture method of mesenchymal stem cells

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Embodiment 1

[0036] Embodiment 1 Serum-free medium preparation:

[0037] The components of the serum-free medium in this embodiment include:

[0038] Non-essential amino acids: 1% by volume

[0039] Glutamine: 2mM

[0040] Hydrocortisone: 50μg / L

[0041] Dexamethasone: 10μg / L

[0042] Polyvinyl alcohol: 2g / L

[0043] Recombinant human insulin: 10mg / L

[0044] Recombinant human transferrin: 5.5mg / L

[0045] Recombinant human epidermal growth factor: 20μg / L

[0046] Recombinant human basic fibroblast growth factor: 20μg / L

[0047] Recombinant human Wnt-3a protein: 20μg / L

[0048] Recombinant human fibronectin: 50μg / L

[0049] Recombinant human laminin: 20μg / L

[0050] L-Glutathione: 4mg / L

[0051] L-Ascorbic acid: 50mg / L

[0052] β-mercaptoethanol: 2.5mg / L

[0053] Ethanolamine: 0.2g / L

[0054] Pluronic F-68: 500mg / L

[0055] Tween 22mg / L

[0056] Cholesterol: 2.2mg / L

[0057] Adenine: 10mg / L

[0058] Sodium selenite: 0.00067mg / L

[0059] DMEM / F12 basal medium: balance. ...

Embodiment 2

[0061] Embodiment 2 Serum-free medium preparation:

[0062] The components of the serum-free medium in this embodiment include:

[0063] Non-essential amino acids: 1% by volume

[0064] Glutamine: 2mM

[0065] Hydrocortisone: 50μg / L

[0066] Dexamethasone: 10μg / L

[0067] Polyvinyl alcohol: 4g / L

[0068] Recombinant human insulin: 10mg / L

[0069] Recombinant human transferrin: 5.5mg / L

[0070] Recombinant human epidermal growth factor: 20μg / L

[0071] Recombinant human basic fibroblast growth factor: 20μg / L

[0072] Recombinant human Wnt-3a protein: 20μg / L

[0073] Recombinant human fibronectin: 50μg / L

[0074] Recombinant human laminin: 20μg / L

[0075] L-Glutathione: 4mg / L

[0076] L-Ascorbic acid: 50mg / L

[0077] β-mercaptoethanol: 2.5mg / L

[0078] Ethanolamine: 0.2g / L

[0079] Pluronic F-68: 500mg / L

[0080] Tween 22mg / L

[0081] Cholesterol: 2.2mg / L

[0082] Adenine: 10mg / L

[0083] Sodium selenite: 0.00067mg / L

[0084] DMEM / F12 basal medium: balance. ...

Embodiment 3

[0086] Embodiment 3 serum-free medium preparation:

[0087] The components of the serum-free medium in this embodiment include:

[0088] Non-essential amino acids: 1% by volume

[0089] Glutamine: 2mM

[0090] Hydrocortisone: 50μg / L

[0091] Dexamethasone: 10μg / L

[0092] Polyvinyl alcohol: 10g / L

[0093] Recombinant human insulin: 10mg / L

[0094] Recombinant human transferrin: 5.5mg / L

[0095] Recombinant human epidermal growth factor: 20μg / L

[0096] Recombinant human basic fibroblast growth factor: 20μg / L

[0097] Recombinant human Wnt-3a protein: 20μg / L

[0098] Recombinant human fibronectin: 50μg / L

[0099] Recombinant human laminin: 20μg / L

[0100] L-Glutathione: 4mg / L

[0101] L-Ascorbic acid: 50mg / L

[0102] β-mercaptoethanol: 2.5mg / L

[0103] Ethanolamine: 0.2g / L

[0104] Pluronic F-68: 500mg / L

[0105] Tween 22mg / L

[0106] Cholesterol: 2.2mg / L

[0107] Adenine: 10mg / L

[0108] Sodium selenite: 0.00067mg / L

[0109] DMEM / F12 basal medium: balance....

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Abstract

The invention relates to the technical field of cell culture, and in particular to a serum-free culture medium, a preparation method thereof and a culture method of mesenchymal stem cells. The culturemedium consists of the following components: non-essential amino acid, glutamine, hydrocortisone, dexamethasone, polyvinyl alcohol, recombinant human insulin, recombinant human transferrin, a recombinant human epidermal growth factor, a recombinant human basic fibroblast growth factor, a recombinant human Wnt-3a protein, a recombinant human fibronectin, recombinant human laminin, L-glutathione, L-ascorbic acid, beta-mercaptoethanol, ethanolamine, Pluronic F-68, Tween 80, cholesterol, adenine, sodium selenite and a basic culture medium. According to the invention, polyvinyl alcohol can replaceserum albumin in a serum-free culture medium in carrier function, so that proliferation capacity of mesenchymal stem cells is effectively improved. Polyvinyl alcohol has more stable performance thanserum albumin, and the problem that proliferation capacity and passage capacity of stem cells are reduced can be effectively solved.

Description

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Claims

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Application Information

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Owner GUANGZHOU SALIAI STEMCELL SCI & TECH CO LTD
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