A kind of magnetic composite microcarrier and its preparation method and application in cell suspension culture
A cell suspension and microcarrier technology, applied in microcarriers, tissue culture, animal cells, etc., can solve the problems of microcarrier collision and cell impact, and achieve the effects of enhanced mechanical strength, uniform particle size distribution, and rich sources of raw materials
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Embodiment 1
[0044] Embodiment 1: the preparation of S-PLLA
[0045]According to the pentaerythritol: lactide molar ratio of 1:300 into the ampoule. Add stannous isooctanoate, add a stirring magnet, blow with nitrogen for 5-6 times, vacuumize, and seal with an alcohol blowtorch. Put it in an oil bath and stir the reaction at 140°C for 24h. Take out the ampoule bottle in the oil bath, take out the material from the bottle, dissolve it in chloroform, slowly add it dropwise in methanol and stir according to the volume ratio of chloroform:methanol is 1:10, then let stand, after the solution is layered, The lower precipitate was taken out and dried in a vacuum oven for 12 h. Precipitate, dry three times and take out to obtain S-PLLA.
[0046] from figure 1 It can be seen from the infrared analysis that lactide and S-PLLA have a wave number of 2997cm -1 There are methyl stretching vibration absorption peaks everywhere, 2936cm -1 、2943cm -1 The methine stretching vibration peak at 1756cm ...
Embodiment 2
[0048] Embodiment 2: the synthesis of ferric oxide nanoparticles
[0049] Weigh a certain amount of Fe according to the ratio of 1.0:1.7 molar ratio 2+ , Fe 3+ Salt, dissolved in 20ml of deionized water, filtered to remove undissolved solids, pour the solution into a three-necked flask, heat to 65°C, add 2mol / L ammonia water to adjust the pH value to 9-10 under the action of high-speed stirring, and continue heating Stir for 0.5h. After the precipitation is complete, pour off the supernatant, wash repeatedly until neutral, put it in an oven to dry for 24h, and collect Fe 3 o 4 Nanoparticles.
[0050] image 3 Shown A, B, C figure is the prepared Fe 3 o 4 TEM image of the nanoparticles, the synthesized Fe was found from image A 3 o 4 The nanoparticles are all granular, and Fe is found in B after magnification 3 o 4 The nanoparticles are spherical, and the diameter of the nanoparticles is found in Figure C to be about 10nm. It can be seen from the figure that the agglo...
Embodiment 3
[0051] Embodiment 3: the preparation of calcium alginate hydrogel
[0052] Different concentrations of SA solution and calcium chloride solution were prepared, and the SA solution was sprayed into the calcium chloride solution under a high-voltage electrostatic spray device to form calcium alginate hydrogel.
[0053] The results show that 0.5-2% (W / V) SA solution has better fluidity, and the viscosity above 2% is higher, which affects the later electrostatic spraying process. Finally, a 1.5% SA solution was used for operation.
[0054] Calcium ion solution uses 4% calcium chloride solution.
[0055] Since the particle size of the microcarrier is required to be between 60-250μm, the voltage needs to be adjusted to a certain value. The high voltage is 20kv, and the low voltage is 7kv, so that microbeads with the same particle size can be ejected.
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