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Human amnion membrane epithelial cell culture medium and application

A technology of epithelial cells and culture medium, applied in the direction of embryonic cells, culture process, tissue culture, etc., can solve the problems of complex components of fetal bovine serum, large differences between batches, disease transmission, etc., to avoid zoonotic diseases and immunity Risk of Reaction, Avoidance of Experimental Uncertainty, Effects of Preserving Differentiation Capacity

Active Publication Date: 2018-08-17
北京科霖恩生物科技有限公司 +1
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AI Technical Summary

Problems solved by technology

However, the composition of fetal bovine serum is complex, and there are large differences between batches. Clinical application has the risk of causing disease transmission between humans and animals and causing immune reactions.
However, the current commercialized epithelial cell serum-free medium is far less effective in expanding hEC than this serum-containing system

Method used

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  • Human amnion membrane epithelial cell culture medium and application
  • Human amnion membrane epithelial cell culture medium and application
  • Human amnion membrane epithelial cell culture medium and application

Examples

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Embodiment 1

[0025] Experiment 1 Isolation, culture and morphological observation of human amniotic membrane pluripotent stem cells (epithelial cells)

[0026] 1. Prepare serum-free medium: prepare serum-free medium as shown in Table 2.

[0027] Table 2. The formula of the serum-free medium that embodiment uses

[0028]

[0029] 2. Isolation of Placental Amniotic Epithelial Cells

[0030] Specimens: Human placentas are taken from fetuses after normal delivery. The use of human specimens complies with the regulations on the application of human specimens in experimental research formulated by the Ethics Committee of the Air Force General Hospital of the Chinese People's Liberation Army.

[0031] Isolation steps: Isolation of human amniotic epithelial cells was carried out according to the method reported in the literature (Murphy S, et al. Amnion Epithelial Cell Isolation and Characterization for Clinical Use. Current Protocol Stem Cell Biology 2010; 13:1E.6.1-1E.6.25). In brief, the a...

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Abstract

The invention provides a human amnion membrane epithelial cell culture medium and application. The culture medium is prepared from (1) an improved MCDB153 basic culture medium, (2) a serum substitution composition, (3) a cell growth factor composition and (4) a micromolecular substance. The culture medium is free of serum or plasma (including animal or human serum or plasma), so that the risk of zoonosis caused by animal serum or plasma in a cell therapy process can be avoided by adoption of the system for human epithelial cell culture. In addition, the culture medium is clear in component, and batch differences caused by use of blood products such as serum, plasma, blood platelets or the like are avoided. The culture medium is suitable for culture of human epithelial cells including amnion membrane epithelial cells and has a promising application prospect in researches of tissue engineering and the like.

Description

technical field [0001] The invention belongs to the field of in vitro culture of cells and tissues, and relates to a medium for culturing human amniotic epithelial cells and its application. The culture medium is used to amplify and cultivate pluripotent stem cells, and is especially suitable for clinical trial research. Background technique [0002] Stem cells refer to cells with multilineage differentiation ability and potentially unlimited self-renewal ability. In the process of embryonic development, the cells that appear in the fertilized egg to the morula stage have the ability to form a complete individual, called totipotent stem cells. As the embryo progresses into the blastocyst stage, a vesicular tissue consisting of the vesicle wall and inner cell mass emerges. The inner cell mass develops into an individual and differentiates into all the tissue cells needed by the individual, called embryonic stem cells, which are pluripotent stem cells. The vesicle wall devel...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/073
CPCC12N5/0605C12N2500/20C12N2500/25C12N2500/32C12N2500/36C12N2500/46C12N2501/11C12N2501/115C12N2501/148C12N2501/315C12N2501/392C12N2501/395C12N2501/855C12N2501/998
Inventor 郭子宽朱笑飞
Owner 北京科霖恩生物科技有限公司
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