New application of thyroxine, method for culturing pluripotent stem cells and culture medium

A technology of pluripotent stem cells and thyroxine, applied in the new application field of drugs, can solve the problems of little research on the influence of cells, and achieve the effect of increasing growth rate and promoting embryonic development.

Pending Publication Date: 2021-07-06
UNIVERSITY OF MACAU
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Problems solved by technology

[0003] In addition, thyroxine plays an important role in embryo implantation and early development in the early stage of embryonic development. Thyroxine has been widely used in the media containing serum, but the effect of thyroxine on cells in these media is rarely seen. have research

Method used

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  • New application of thyroxine, method for culturing pluripotent stem cells and culture medium
  • New application of thyroxine, method for culturing pluripotent stem cells and culture medium
  • New application of thyroxine, method for culturing pluripotent stem cells and culture medium

Examples

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Effect test

experiment example 1

[0044] Thyroxine T3 promotes the growth rate of human pluripotent stem cells in suspension culture

[0045] Method: Suspension culture of human pluripotent stem cells takes advantage of the space advantage of the medium to free the cells from the plane limitation of adherent culture, increase the utilization rate of the medium, and contribute to the large-scale industrial culture of pluripotent stem cells. However, in the practice of suspension culture, it is found that the growth rate of cells is greatly slowed down compared with that of adherent culture. In this experiment, taking the small-scale culture in the laboratory as an example, it is proved that thyroxine T3 plays a role in the suspension culture of human pluripotent stem cells. effect. The specific experimental conditions are 5% CO2, constant temperature culture at 37°C, and the experimental basal medium is E8 medium (components: DMEM / F12, L-ascorbic acid-2-phosphate magnesium (64mg / l), sodium selenium (14μg / l), ...

experiment example 2

[0048] Thyroxine T3 promotes the stability of human pluripotent stem cells in adherent culture

[0049] Method: With reference to the concentration of thyroxine T3 in Experimental Example 1, E8 medium containing 500nM thyroxine T3 was added during cell adherent culture, and the cells were incubated at 37°C in 5% CO 2 cultured in an incubator. Then, referring to the previous thyroid concentration range, the inhibitory effect of 5-500nM thyroxine on apoptosis molecules in high-density cultured stem cells was tested. Subsequently, the optimized E8 medium containing 500nM thyroxine was used to detect the effects of cell subculture survival efficiency after high-density culture, normal density cloning efficiency, cell subculture stability and pluripotency.

[0050] For test results, see figure 2 ,in, figure 2 Middle A is the result of immunofluorescence experiment of apoptosis molecule caspase3 / 7, figure 2 In B, the flow cytometry diagram of the effect of different concentra...

experiment example 3

[0053] Thyroxine T3 Promotes Metabolism of Human Pluripotent Stem Cells

[0054] Methods: (1) Human pluripotent stem cells were cultured in E8 medium containing 500nM thyroxine T3 for two days, and the mitochondrial stress test of Seahorse cell metabolism analysis platform was used to detect the effect of thyroxine T3 on the mitochondria of human pluripotent stem cells.

[0055] (2) Human pluripotent stem cells were cultured in E8 medium containing 500nM thyroxine T3 for two days, and the glycolysis test of Seahorse cell metabolism analysis platform was used to detect the effect of thyroxine T3 on glycolysis of human pluripotent stem cells.

[0056] For test results, see image 3 ,in image 3 Middle A is the result of mitochondrial stress test, image 3 Middle B is the graph of glycolysis test results.

[0057] according to image 3 In A, it can be seen that thyroxine T3 can increase basal respiration (Basal) and ATP production, thereby increasing mitochondrial activity. ...

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Abstract

The invention relates to the technical field of new application of medicines, in particular to new application of thyroxine, a method for culturing pluripotent stem cells and a culture medium. The invention relates to application of thyroxine in preparation of medicines for treating embryonic growth retardation or development insufficiency. The thyroxine can promote embryonic development, then can be used for preparing the medicines for treating delayed embryonic development or embryonic insufficiency, can further be used for improving the growth rate, stability, metabolic level, differentiation and the like of the pluripotent stem cells, and can be further used for preparing a reagent for achieving the effects.

Description

technical field [0001] The present invention relates to the technical field of new application of medicine, in particular, relates to the new application of thyroxine, the method and medium for cultivating pluripotent stem cells. Background technique [0002] Human pluripotent stem cells (hPSCs) have the potential to differentiate into all cell types, and play an important role in the study of human embryonic development, disease model research, drug screening and cell therapy. Traditional human pluripotent stem cell culture techniques include small-scale plate adherent culture and large-scale suspension culture. For human pluripotent stem cell plate adherent culture, the cell expansion speed is fast, but due to surface limitations, cell growth and survival are inhibited under high-density culture, and it is not suitable for large-scale industrial cell culture. Large-scale cell culture often uses cell suspension Cultivate technology. In the process of cell suspension cultu...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K31/198A61P43/00C12N5/074
CPCA61K31/198C12N5/0696A61P43/00C12N2501/395C12N2500/25C12N2500/38C12N2501/115C12N2501/15
Inventor 陈国凯邓春浩
Owner UNIVERSITY OF MACAU
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