Composition, culture medium supplement, stem cell culture medium and stem cell culture method

A technology for culturing and cultivating stem cells, applied in the field of composition, stem cell culture medium and culturing, can solve the problems of cell vacuolization, decreased expansion ability, cell rupture and death, etc., and achieve cell vacuolation inhibition and cell senescence alleviation , the effect of low cost

Active Publication Date: 2021-08-06
BIOISLAND LAB +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] When ADSCs are cultured in a serum-free culture system, the problems they face are more severe than other types of MSCs, because ADSCs are adult stem cells, and their in vitro expansion ability itself is relatively limited. If ADSCs are cultured in a serum-free medium in vitro, the expansion ability There will be a significant decrease, and the phenomenon of cell vacuolation is very obvious after continuous passage. As the number of passages increases, the cytoplasmic vacuoles age rapidly, and as the vacuoles become larger, the cells eventually rupture. die

Method used

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  • Composition, culture medium supplement, stem cell culture medium and stem cell culture method
  • Composition, culture medium supplement, stem cell culture medium and stem cell culture method
  • Composition, culture medium supplement, stem cell culture medium and stem cell culture method

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preparation example Construction

[0055] An embodiment of the present invention also provides a method for preparing a stem cell culture medium as described above, the preparation method comprising the following steps: mixing the composition or medium supplement and the basal medium.

[0056] The embodiment of the present invention relates to the application of the above composition, the medium supplement or the stem cell culture medium in stem cell culture.

[0057] In one example, the composition, the medium supplement and the stem cell culture medium inhibit stem cell senescence and / or improve stem cell proliferation efficiency.

[0058] In one example, the composition, the medium supplement, and the stem cell culture medium prevent vacuolization of stem cell subcultures.

[0059] The embodiment of the present invention also provides a stem cell culture method, the culture method comprising the following steps:

[0060] Stem cells were inoculated into the stem cell culture medium as described above, and cu...

Embodiment 1

[0067] This embodiment provides a culture medium for adipose-derived mesenchymal stem cells and a preparation method thereof. Specifically, the following steps are involved:

[0068] (1) Take L-ascorbic acid-2-phosphate trisodium salt, catalase, coenzyme Q10, rapamycin, apocynum and non-essential amino acid solution, and prepare a solution containing 25mg / L L-ascorbic acid-2-phosphate Culture of trisodium salt, 1 mg / L catalase, 1 μM coenzyme Q10, 1 μM rapamycin, 1 μM apocynin and 1% (v / v) non-essential amino acid solution (Gibco11140-050 non-essential amino acid solution) Base supplement, 0.22μm filter membrane filter sterilization. The medium supplement was used as a storage solution (1000×), and stored at -20°C to -80°C for future use.

[0069] The preparation step of the storage solution includes: dissolving each component according to its respective characteristics, and then using DPBS as a solvent, mixing and preparing the storage solution (1000×) with the above concent...

Embodiment 2

[0072] This embodiment provides a culture medium for adipose-derived mesenchymal stem cells and a preparation method thereof. Specifically, the following steps are involved:

[0073] (1) Take L-ascorbic acid-2-phosphate trisodium salt, catalase, coenzyme Q10, rapamycin, apocynum and non-essential amino acid solution, and prepare a solution containing 50mg / L L-ascorbic acid-2-phosphate Culture of trisodium salt, 10 mg / L catalase, 10 μM coenzyme Q10, 10 μM rapamycin, 10 μM apocynin and 1% (v / v) non-essential amino acid solution (Gibco11140-050 non-essential amino acid solution) Base supplement, 0.22μm filter membrane filter sterilization. The medium supplement was used as a storage solution (1000×), and stored at -20°C to -80°C for future use. Refer to Example 1 for the preparation steps of the stock solution (1000×).

[0074] (2) Take the above-mentioned medium supplement and add it to the serum-free medium UltraCULTURE Serum-free Medium of LONZA Company and Guangdong Guoke ...

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Abstract

The invention relates to a composition, a culture medium supplement, a stem cell culture medium and a stem cell culture method. The composition is prepared from L-ascorbic acid-2-trisodium phosphate, catalase, coenzyme Q10, rapamycin and apocynin. According to the invention, L-ascorbic acid-2-trisodium phosphate, catalase, coenzyme Q10, rapamycin and apocynin are selected to be combined to form a composition with a specific formula, and the composition and non-essential amino acids are added into a serum-free culture medium to form a specific serum-free culture system. When the serum-free culture system is adopted to perform subculture on ADSCs, the cell proliferation rate can be higher than that of a serum-containing culture system. Meanwhile, in subculture, cell vacuolation is effectively inhibited, cell senescence is relieved, and the effect can reach the level equivalent to that of a serum culture medium.

Description

technical field [0001] The invention relates to the technical field of cell culture, in particular to a composition, a culture medium supplement, a stem cell culture medium and a culture method. Background technique [0002] Mesenchymal stem cells (MSCs) are derived from the mesoderm in the early stage of development. tissue, fat, fatty blood and placenta and other tissues. MSCs have high self-renewal ability and multi-directional differentiation potential, and can be cultured and expanded in vitro. They can not only support the growth of hematopoietic stem cells, but also have the function of immune regulation; under different induction conditions, they can differentiate into bone, cartilage, and muscle in vitro. , nerve, myocardium, endothelium and fat, etc., still have multi-directional differentiation potential after continuous subculture and cryopreservation, and can be used as ideal seed cells for the repair of tissue and organ damage caused by aging and disease. The...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0775
CPCC12N5/0662C12N5/0667C12N2500/32C12N2500/38C12N2500/90C12N2500/30C12N2501/71C12N2501/999
Inventor 陈东煌陈海佳戚康艺姜交华李学家
Owner BIOISLAND LAB
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