Method for Suspension Culture Cells to Produce Clematis Polysaccharides
A technology of suspension culture cells and clematis, which is applied in fermentation and other directions, can solve the problems of long production cycle, low biomass growth rate, low yield of target products, etc., and achieve improved product yield, uniform nutrition and dissolved oxygen supply , the effect of shortening the cell culture cycle
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specific Embodiment 1
[0017] 1. Training stage
[0018] (1) Single cell preparation
[0019] Take clematis capsules, sterilize them according to the requirements of tissue culture technology, and carry out aseptic sowing. The sowing medium is an improved 1 / 2MS solid medium. Take them out after they germinate into protocorms. Choose protocorms with bright color, large individuals and vigorous vitality. For bulbs, separate and prepare single cells by enzymatic hydrolysis, filter with a microporous filter with a pore size of 80 microns, separate the filtrate through a low-speed centrifuge, and collect the precipitated cells;
[0020] (2) Single cell line culture
[0021] Inoculate the collected single cells into a baffle shake flask containing improved 1 / 2MS liquid medium, the inoculum amount of fresh cells is 25±2g / L, the light intensity is 3000lux, the color temperature is 5000K, shake culture, adjust the pH value 5.6±2, temperature 22±2°C, when the cell density reaches 10000 cells / mL, the culture...
specific Embodiment 2
[0031] 1. Training stage
[0032] (1) Single cell preparation
[0033] Take clematis capsules, sterilize them according to the requirements of tissue culture technology, and carry out aseptic sowing. The sowing medium is an improved 1 / 2MS solid medium. Take them out after they germinate into protocorms. Choose protocorms with bright color, large individuals and vigorous vitality. For bulbs, single cells were separated by enzymatic hydrolysis, filtered through a microporous filter with a pore size of 60 microns, the filtrate was separated by a low-speed centrifuge, and the precipitated cells were collected;
[0034] (2) Single cell line culture
[0035] Inoculate the collected single cells into a baffle shake flask containing improved 1 / 2MS liquid medium, the inoculum amount of fresh cells is 25±2g / L, the light intensity is 2000lux, the color temperature is 6000K, shake culture, adjust the pH value 5.6±2, temperature 22±2°C, when the cell density reaches 20,000 / mL, it can be ...
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