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Serum-free complete medium and use thereof

A complete medium, serum-free culture technology, applied in the field of serum-free complete medium, can solve the problem of complex medium components, achieve the effect of maintaining "stemness" and multi-directional differentiation ability, and avoiding pollution

Inactive Publication Date: 2018-11-06
安徽瑞达健康产业有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

This patented technology allows for easy preparation of various ingredients needed during culture process without causing any harm or damage from bacteria that may be introduced into the tissue being grown. It has been found that knockout SR can lead towards more efficient cell growth compared with other methods like plating on slides containing certain chemical compounds (KSR) which could potentially cause further damaging agents such as germicidal drugs). Additionally, it provides an effective way to study dentinogenesis processes through studying periodontitis patients' teeth ex vivo.

Problems solved by technology

The technical problem addressed in this patents relates to developing an improved method for producing mammals containing both somatic stem cells like placenta and spinal cord regenerating stem cells without causing any side effects associated therewith those previously described methods.

Method used

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  • Serum-free complete medium and use thereof
  • Serum-free complete medium and use thereof
  • Serum-free complete medium and use thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0052] Example 1: Serum-free culture of deciduous dental pulp mesenchymal stem cells

[0053] Serum-free complete medium I: DMEM basal medium, 2mM / L L-glutamine, 2% Ultroser G solution, mix well, store in refrigerator at 4°C.

[0054] Serum-free complete medium II: containing DMEM basal medium, 2mM / L L-glutamine, 10% knockout SR serum replacement, 10ng / ml EGF, 10ng / ml bFGF.

[0055] Training method:

[0056] 1) Freshly exfoliated deciduous teeth were separated by Masako Miura et al. After digestion with type I collagenase, they were filtered and centrifuged to obtain mesenchymal stem cells for primary culture. The primary culture used serum-free complete medium I and serum-free medium complete ⅡCultivation in the medium after equal volume mixing, the culture period is 7-14 days;

[0057] 2) Discard the original cell culture medium in the T75 square flask cultured with P0 primary tooth mesenchymal stem cells, add 7.5ml DPBS to wash once;

[0058] 3) Add 0.8ml of 0.25% trypsi...

Embodiment 2

[0068] Example 2 Serum-free culture of deciduous dental pulp mesenchymal stem cells

[0069] Mix medium I and medium II to prepare medium III: DMEM basal medium, 2mM / L L-glutamine, 1% Ultroser G solution, 5% KnockOut SR serum replacement, 10ng / ml EGF, 10ng / ml bFGF and mix well, store in refrigerator at 4°C.

[0070] Training method:

[0071] 1) The freshly exfoliated deciduous teeth are separated by Masako Miura et al. After digesting with type I collagenase, they are filtered and centrifuged to obtain mesenchymal stem cells for primary culture. The primary culture is cultured with medium III, and the culture period is 7-14 days. sky;

[0072] 2) Discard the original cell culture medium in the T75 square flask cultured with P0 primary tooth mesenchymal stem cells, add 7.5ml DPBS to wash once;

[0073] 3) Add 0.8ml of 0.25% trypsin to digest for 4 minutes and resuspend the cells with serum-free complete medium III;

[0074] 4) Transfer the cell suspension to a sterile cent...

Embodiment 3

[0079] Example 3: Serum-free culture of deciduous dental pulp mesenchymal stem cells

[0080] Serum-free complete medium I: DMEM basal medium, 2mM / L L-glutamine, 4% Ultroser G solution, mix well, store in refrigerator at 4°C.

[0081] Serum-free complete medium II: containing DMEM basal medium, 2mM / L L-glutamine, 10% KnockOut SR serum replacement, 10ng / ml EGF, 10ng / ml bFGF.

[0082] Training method:

[0083] 1) Freshly exfoliated deciduous teeth were separated by Masako Miura et al. After digestion with type I collagenase, they were filtered and centrifuged to obtain mesenchymal stem cells for primary culture. The medium was 4ml serum-free complete medium II and 1ml serum-free complete culture The mixed medium of base Ⅰ, the culture period is 7-14 days;

[0084] 2) Discard the original cell culture medium in the T75 square flask cultured with P0 primary tooth mesenchymal stem cells, add 5ml DPBS to wash once;

[0085] 3) Add 0.8ml 0.25% trypsin to digest for 4 minutes, then...

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Abstract

The invention provides a serum-free complete medium and a use thereof. The serum-free complete medium comprises two mediums which are orderly used or used together. The medium I contains L-glutamine with the concentration of 2mM/L and 1-4% of a serum substitute for effectively promoting cell adhesion and extension based on a basic medium. The medium II contains L-glutamine with the concentration of 2mM/L and 7-10% of a serum substitute for culturing ESC and iPSC based on a basic medium. The invention also provides the use of the serum-free complete medium in culture of mesenchymal stem cells.The serum-free complete medium utilizes simple medium ingredients and is easy to prepare. The mesenchymal stem cell culture method is conducive to in-vitro culture of human deciduous teeth mesenchymalstem cells and keeps the dryness and multi-directional differentiation ability of stem cells.

Description

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Claims

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Application Information

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Owner 安徽瑞达健康产业有限公司
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