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Culture medium supplement, culture medium supplement composition, culture medium and culture method thereof

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 stemness of pluripotent stem cells, and achieve the effect of avoiding unstable medium performance and maintaining survival and dryness

Active Publication Date: 2021-02-12
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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  • Culture medium supplement, culture medium supplement composition, culture medium and culture method thereof
  • Culture medium supplement, culture medium supplement composition, culture medium and culture method thereof
  • Culture medium supplement, culture medium supplement composition, culture medium and culture method thereof

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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 kaempferol and 1 μM SRI-011381hydrochloride.

[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-011381hydrochloride.

[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 culture medium supplement, a culture medium supplement composition, a culture medium and a culture method thereof. According to the present invention, kaempferitrin and SRI011381 hydrochlolide are combined to form the specific culture medium supplement, such that the survival and the dryness of pluripotent stem cells can be effectively maintained when the pluripotent stem cells are cultured by using the culture medium containing the culture medium supplement; moreover, when the culture medium containing the culture medium supplement is used for culturing the pluripotent stem cells, compared with the traditional pluripotent stem cell culture medium containing human recombinant insulin and human recombinant TGF-beta1 protein, the problems of unstable culture mediumperformance and the like caused by inactivation of various recombinant proteins are avoided to a certain extent.

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 Applications(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
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