A serum-free and feeder-free culture medium for embryonic stem cells or pluripotent stem cells and its application

A technology for embryonic stem cells and pluripotent stem cells, applied in the field of embryonic stem cells or pluripotent stem cell culture medium, can solve the problems of increasing the cost of serum-free medium, reducing the bioavailability of pluripotency, and instability affecting the quality of hESCs, etc. Achieve the effect of reducing preparation cost, maintaining stable quality and reducing aggregation phenomenon

Active Publication Date: 2021-07-16
BIOISLAND LAB +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, bFGF signaling is dose-dependent and high levels of bFGF may be required to meet specific signaling thresholds or to prevent inhibitory effects such as protein degradation
At the same time, bFGF will aggregate at 37°C, thereby reducing the bioavailability to maintain pluripotency
Therefore, as an important signaling factor, bFGF’s high concentration increases the cost of serum-free medium, and its instability will affect the quality of hESCs. Researchers engaged in hESCs research have to face the dilemma of cost and quality

Method used

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  • A serum-free and feeder-free culture medium for embryonic stem cells or pluripotent stem cells and its application
  • A serum-free and feeder-free culture medium for embryonic stem cells or pluripotent stem cells and its application
  • A serum-free and feeder-free culture medium for embryonic stem cells or pluripotent stem cells and its application

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Experimental program
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Embodiment 1

[0051] This embodiment provides a serum-free feeder-free culture medium for embryonic stem cells or pluripotent stem cells, the formula of which is shown in the following table:

[0052]

[0053]

[0054] The preparation method is as follows: dissolving each additive component in the culture medium in the above table according to their respective dissolution characteristics, and filtering and sterilizing through a filter membrane. Under sterile conditions at 20°C, add each component to the DMEM / F12 basal medium one by one, pipette and mix well, adjust the osmotic pressure to 340mOSM / kg with sodium chloride, store the prepared medium at 4°C, and store it within two weeks efficient.

Embodiment 2

[0056] This embodiment provides a serum-free feeder-free culture medium for embryonic stem cells or pluripotent stem cells, the formula of which is shown in the following table:

[0057]

[0058]

[0059] Its preparation method refers to embodiment 1.

Embodiment 3

[0061] This embodiment provides a serum-free feeder-free culture medium for embryonic stem cells or pluripotent stem cells, the formula of which is shown in the following table:

[0062]

[0063] Its preparation method refers to embodiment 1.

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Abstract

The present invention relates to a serum-free feeder-free embryonic stem cell or pluripotent stem cell culture medium and applications thereof. The serum-free feeder-free embryonic stem cell or pluripotent stem cell culture medium includes a basal medium and additives, and the additives include Sodium heparin, pipecolic acid, gamma-aminobutyric acid, and lithium ion salts. It is a completely xeno-free culture system that will not cause external contamination; the concentration of bFGF in the medium can be significantly reduced, thus significantly reducing the cost of medium preparation and effectively reducing The aggregation phenomenon of bFGF is eliminated, the stability is improved, the quality of cultured embryonic stem cells or pluripotent stem cells is maintained stable, and the undifferentiated state and pluripotency of embryonic stem cells or pluripotent stem cells can be maintained for a long time; in addition, pipecolic acid, γ ‑GABA and lithium ion salt have an unpredictable synergistic effect in promoting the survival, proliferation and maintenance of continuous passage of embryonic stem cells or pluripotent stem cells.

Description

technical field [0001] The invention belongs to the technical field of cell culture, and relates to a medium for embryonic stem cells or pluripotent stem cells and its application, in particular to a serum-free and feeder-free medium for embryonic stem cells or pluripotent stem cells and its application, in particular to a stable Embryonic stem cell or pluripotent stem cell culture medium with low protein content and application thereof. Background technique [0002] Human embryonic stem cells (hESCs) are pluripotent cells derived from the inner cell mass of blastocysts fertilized in vitro. hESCs have the ability to differentiate into ectoderm, endoderm and mesoderm, such as cardiomyocytes, neurons, glial cells, hepatocytes, islet cells, chondrocytes, skeletal muscle cells, adipocytes and endothelial cells. [0003] The traditional hESCs culture method needs to place hESCs on mitomycin-treated feeder cells (such as embryonic fibroblasts, MEFs). Although this method can main...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/0735C12N5/074
CPCC12N5/0606C12N5/0696C12N2500/05C12N2500/12C12N2500/14C12N2500/22C12N2500/24C12N2500/30C12N2500/38C12N2500/60C12N2500/90C12N2501/115C12N2501/15C12N2501/415C12N2501/845C12N2501/91C12N2501/999
Inventor 陈东煌陈海佳姜交华张兆清王小燕李学家
Owner BIOISLAND LAB
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