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Media Supplement, Media Supplement Composition, Media, and Culture Method

A technology for supplementing compositions and culture methods, which is applied in the field of medium supplements, can solve problems such as inability to effectively maintain the survival and dryness of pluripotent stem cells, and achieve the effect of avoiding unstable medium performance and maintaining survival and dryness

Active Publication Date: 2022-03-01
BIOISLAND LAB +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] However, traditional pluripotent stem cell culture medium as mentioned above cannot effectively maintain the viability and stemness of pluripotent stem cells

Method used

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  • Media Supplement, Media Supplement Composition, Media, and Culture Method
  • Media Supplement, Media Supplement Composition, Media, and Culture Method
  • Media Supplement, Media Supplement Composition, Media, and Culture Method

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preparation example Construction

[0048] An embodiment of the present invention provides a method for preparing the cell culture medium as described above, the preparation method comprising the following steps:

[0049] The components contained in the above-mentioned medium supplement or the components contained in the above-mentioned medium supplement composition are dissolved and added to the basal medium.

[0050] As mentioned above, the osmotic pressure of the culture medium provided by the embodiments of the present invention can be adjusted to an appropriate range according to actual needs, such as 290mOSM / kg-340mOSM / kg.

[0051] An embodiment of the present invention provides a method for culturing pluripotent stem cells, the culturing method comprising the following steps:

[0052] The pluripotent stem cells are inoculated in the above-mentioned cell culture medium and cultured.

[0053] It can be understood that, the embodiment of the present invention does not specifically limit the specific type of...

Embodiment 1

[0057] This embodiment provides a cell culture medium and a preparation method thereof.

[0058] The formulation of the medium is shown in Table 1, including: 500mg / L of NaHCO 3 , 13 μg / L apotransferrin, 64 mg / L L-ascorbic acid, 15 μg / L sodium selenite, 35 μg / L recombinant human basic fibroblast growth factor, 6.7 mg / L thiamine hydrochloride Glutathione, 7 mg / L reduced glutathione, 10 μM poloxamer 188, 10 μM kaempferin and 1 μM SRI-011381 hydrochloride.

[0059] The preparation method of the culture medium includes: dissolving the various additive components shown in the culture medium in the table according to their respective solubility characteristics, and after filter sterilization; In the F12 basal medium, pipette and mix well, adjust the osmotic pressure to 310mOSM / kg with sodium chloride; store the prepared medium at 4°C.

Embodiment 2

[0061] This embodiment provides a cell culture medium and a preparation method thereof.

[0062] See Table 1 for the formulation of the medium, including: 100mg / L of NaHCO 3 , 50 μg / L apotransferrin, 30 mg / L L-ascorbic acid, 1 μg / L sodium selenite, 10 μg / L recombinant human basic fibroblast growth factor, 1 mg / L thiamine hydrochloride , 10 mg / L reduced glutathione, 5 μM poloxamer 188, 1 μM kaempferin and 0.1 μM SRI-011381 hydrochloride.

[0063] The preparation method of the culture medium includes: dissolving the various additive components shown in the culture medium in the table according to their respective solubility characteristics, and after filter sterilization; In the F12 basal medium, pipette and mix well, adjust the osmotic pressure to 310mOSM / kg with sodium chloride; store the prepared medium at 4°C.

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Abstract

The invention relates to a medium supplement, a medium supplement composition, a medium and a cultivation method. In the present invention, the combination of kaempferin and SRI‑011381 hydrochloride forms a specific medium supplement, and when the medium containing the medium supplement is used to cultivate pluripotent stem cells, it helps to effectively maintain the survival and stemness of pluripotent stem cells . And, when cultivating pluripotent stem cells with the medium containing this medium supplement, compared with the traditional pluripotent stem cell medium containing such as human recombinant insulin, human recombinant TGF-β1 protein, to a certain extent, avoiding various recombinant proteins Problems such as unstable medium performance caused by inactivation.

Description

technical field [0001] The invention relates to the technical field of cell culture, in particular to a medium supplement, a medium supplement composition, a medium and a culture method. Background technique [0002] Pluripotent stem cells (PSCs), including embryonic stem cells (ESCs) and induced pluripotent stem cells (iPSCs), have the ability to differentiate into various types of cells such as cardiomyocytes, neurons, and liver cells. It is not easy to maintain the pluripotency and self-renewal of pluripotent stem cells, and often requires a variety of cytokines and compounds to achieve a certain effect. Correspondingly, a variety of media containing these cytokines and compounds have emerged. [0003] Traditional medium for maintaining the viability and stemness of pluripotent stem cells, such as a serum-free medium made of NaHCO 3 , Sodium Selenite, Iron EDTA, L-Ascorbic Acid, Zinc Sulfate, Recombinant Human Basic Fibroblast Growth Factor, Recombinant Human Transformin...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/0735C12N5/074
CPCC12N5/0606C12N5/0696C12N2500/30C12N2501/999
Inventor 陈东煌陈海佳张兆清姜交华王小燕李学家
Owner BIOISLAND LAB