Anti-aging or antioxidant composition containing plant stem cell line derived from cambium of panax ginseng
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example 1
Preparation of Cell Line Derived From Cambium of Wild Ginseng
[0052]1-1: Preparation of Plant Material
[0053]“A” of FIG. 1(a) shows the typical feature of wild ginseng used in the present invention. In order to use the main root of purchased wild ginseng, the main root was purchased, and then washed with running water to remove earth or other contaminants from the surface thereof, and the surface of the main root was washed with a liquid detergent. Then, the main root was allowed to stand under running water. The washed root tissue was placed in a sterilized flask in a clean bench and disinfected with 70% ethanol for a time ranging from about 30 seconds to about 1 minute. Then, it was rinsed with sterile water and treated with a disinfectant solution containing 1-1.5% sodium hypochlorite (Junsei, Japan) for about 5-15 minutes. At this time, in order to allow the disinfectant solution to effectively penetrate into the tissue, several drops of TWEEN 20 (polyoxyethylenesorbitan monolaura...
example 2
Preparation of Extract of Wild Ginseng Cambium-Derived Cell Line
[0076]Active substances were extracted in the following manner from three cell lines prepared in Example 1:
[0077](1) Preparation of DMSO (dimethyl sulfoxide) Extract
[0078](i) 500 g of each of the cell line, from which the medium has been removed, and the freeze-dried cell line, was dissolved in 500 ml of DMSO with stirring at 50° C. for 6 hours.
[0079](ii) After completion of the dissolution, the cell solution was centrifuged at 3,000 g for 10 minutes, and the supernatant was collected, thus obtaining a DMSO-soluble substance.
[0080](iii) The obtained DMSO-soluble substance was concentrated using a rotary vacuum concentrator.
[0081](iv) The concentrated sample was dried using a freeze dryer, thereby obtaining a DMSO extract.
[0082](2) Preparation of Distilled Water Extract, Methanol Extract and Acetone Extract
[0083](i) 500 g of each of the cell line, from which the culture medium has been removed, and the freeze-dried cell ...
example 3
Examination of Cytotoxicity
[0096]In order to determine the concentration of sample to be used in tests, a cytotoxicity test was performed in the following manner.
[0097]NHF (normal human fibroblast) cells used in the test were isolated from the fetal foreskin and cultured. A medium used in the cell culture was prepared by adding 10% fetal bovine serum (FBS, Hyclone, Logan, Utah, USA), inactivated by heating at 56° C. for 30 minutes, 100 unit / ml penicillin, 100 μg / ml streptomycin and 300 μg / ml-glutamine, to DMEM medium (Invitrogen Gibco life tech. Vienna, Austria). The cells were cultured in the above medium in a 5% CO2 incubator at 37° C. at a relative humidity of 95%, and subcultured at 3 to 4-day intervals immediately before the cells would be fused with each other. The NHF cells were used in the experiments of this Example and Examples 4 and 5 below.
[0098]The NHF(p5) cells (normal human fibroblast cells) were dispensed in 96-well plates at a density of 5,000 cells / well, and after ...
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