Extraction preparation method of star-of-Bethlehem stem cell cambium and anticancer composition
A technology of stem cell line and evergreen, applied in plant cells, food preparation, food science, etc., can solve the problem of hard to find raw materials
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Embodiment 1
[0100] Embodiment 1: the preparation of isolated material
[0101] Prefer the evergreen corm that has grown for 4 years, wash the attachments and soil and other impurities on the surface of the corm with water, put it into a mixed solution of 110mg, L-ascorbic acid and 1% L benomyl for 10 minutes, and then use sterile magnetization Wash with water for 10 minutes, sterilize in 75% ethanol solution for 1 minute, put in 30% hydrogen peroxide solution for 20 minutes, and then rinse with sterile distilled water for 10 minutes.
Embodiment 2
[0102] Embodiment 2: the preparation method of solid medium
[0103] (1) Potassium nitrate 2510mg, ammonium sulfate 136mg, magnesium sulfate 123mg, manganese sulfate 10mg, zinc sulfate 2.5mg, copper sulfate 0.026mg, calcium chloride 114mg, potassium iodide 0.7mg, cobalt chloride 0.026mg, sodium dihydrogen phosphate 131mg, Boric acid 2.8mg, sodium molybdate 0.26mg, iron sodium edetate 37mg.
[0104] Inositol 210mg, Thiamine 21mg, Niacin 2.2mg, Pyridoxine 2.2mg, L-Ascorbic Acid 52mg, Citric Acid 74mg. L-aspartic acid 135mg, L-arginine 176mg, glycine 76mg, proline 116mg, α-naphthaleneacetic acid 2.2mg, sucrose 10.100mg, activated carbon 200mg, agar 3.100mg, magnetized water 0.63 liters.
[0105] (2) Dilute the nutrients of inorganic salts with 20ml of magnetized water, combine vitamins into 50ml of sterile magnetized water for dilution, combine amino acids into 50ml of sterile magnetized water for dilution, and put auxin into 10ml of sterile Dilute in magnetized water. Then in...
Embodiment 3
[0106] Example 3: Induction of isolated materials and separation and culture of cambium.
[0107] (1) Cut the sterilized evergreen corm into 4 sections with an inoculation knife, remove the outermost layer, take the second and third inner cladding layers, cut them into small cubes of 1-2cm, and transfer them to the culture medium , cultured in a controlled dark room in the dark for 28 days at a temperature of 25±1°C. After culturing for 28 days, gently transfer the cambium cell population into another flask of the same medium with an inoculation spoon or inoculation shovel, and then enlarge and cultivate it for 23 days.
[0108] (2) Then transfer the cell line population of its amplified culture into liquid medium, and cultivate in a dark room with a rotary shaker at 100 rpm at 25±1° C., and the passage time is 20 days.
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