Cell three-dimensional culture scaffold, preparation method and application thereof
A three-dimensional culture and cell technology, applied in the fields of biological tissue engineering and biomedicine, can solve the problems of poor cell infiltration effect, lack of precise control of freeze-drying and quenching process, lack of precise control of scaffold shape and parameter pore size, etc.
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Embodiment 1
[0118] Example 1: Preparation of cell three-dimensional culture scaffold (porous scaffold)
[0119] 1. Solution preparation
[0120] 1. First use pure acetic acid and deionized water to prepare a 0.1M / L acetic acid solution (that is, a concentration of 0.6% v / v) in a sterilized container, and then filter it with a 0.22 μm filter membrane and store it in a closed sterile container (not Store in a 4°C refrigerator when in use);
[0121] 2. Use 0.1M / L acetic acid solution to dissolve chitosan (analytically pure, purified from shrimp and crab shells in nature, medium molecular weight range, such as Sigma417963) in a sterilized container, according to the concentration of 2% w / v chitosan The solution is prepared and the top of the container is closed with a cap. Place the solution to be dissolved in a 55°C water bath beaker (the level of the water bath must be higher than the level of the chitosan solution), and use a heatable magnetic stirrer to keep the temperature of the wat...
Embodiment 2
[0141] Embodiment 2: cell culture experiment
[0142] 1. Purpose of the experiment
[0143] Evaluate the effect of chitosan-collagen on cell growth of the three-dimensional scaffold material (50 μl scaffold and 75 μl scaffold) prepared in Example 1, method: cell line HepG2 and primary cell adipose stem cells (cell line HepG2 is purchased from ATCC, adipose stem cell Commercially available or prepared according to conventional methods in the art) were inoculated on the scaffold material, and cultured in vitro for a week, and the number of cells in the scaffold was detected by CCK-8 at different time points.
[0144] 2. Experimental materials and equipment
[0145] As shown in Table 2 and Table 3 below.
[0146] Table 2: Experimental materials
[0147]
[0148]
[0149] Table 3: Experimental equipment
[0150] name
Manufacturer
model
Ultra low temperature centrifuge
Thermo
LEGEND Mach1.6R
STIK
...
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