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.
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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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