Cryogenic freezing method of scaffolds for tissue repair
A low-temperature freezing, tissue repair technology, used in tissue culture, bone/connective tissue cells, biochemical equipment and methods, etc.
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[0033] The preparation method of the gallic acid grafted chitosan scaffold of the present invention is as follows:
[0034] 28 mmol of gallic acid and 2.8 mmol of EDC were first dissolved in 40 mL of 70% ethanol and 2.8 mmol of NHS was added to the solution. The resulting solution was stirred in an ice bath, then 1.5 g of chitosan dispersed in 110 mL of 70% ethanol was added after 1 hour. The solution was further stirred in an ice bath for 30 minutes and finally at room temperature for 24 hours. The precipitate was collected by filtration and washed with ethanol. After dialysis against deionized water for 3 days to remove possible residual reagents, gallic acid-grafted chitosan was obtained by vacuum freeze-drying. Porous gallic acid-grafted chitosan scaffolds were prepared by utilizing the physical properties of gallic acid-grafted chitosan being dissolved in an acidic solution and insoluble in an alkaline condition. First, 1 g of gallic acid grafted chitosan was dissolved...
Embodiment 1
[0044] For the survival rate of mesenchymal stem cells in tissue engineering structures, see figure 1 shown.
[0045] The proliferative ability of mesenchymal stem cells in tissue engineered structures see image 3 shown.
[0046] Colony-forming ability of mesenchymal stem cells in tissue engineered structures see Figure 4 shown, the three differentiation capabilities of mesenchymal stem cells in tissue-engineered structures are shown in Figure 5 shown.
Embodiment 2
[0048] The tissue engineered structures were cryopreserved in 10% DMSO-containing fetal bovine serum (90% FBS) cryopreservation tubes. The cryovials were then sequentially stored at 4°C for 30 minutes, -20°C for 60 minutes, -80°C overnight, and finally stored under liquid nitrogen for 30 days.
[0049] For the survival rate of mesenchymal stem cells in tissue engineering structures after cryopreservation, see figure 1 shown. For the survival rate of mesenchymal stem cells in tissue engineering structures after cryopreservation, see figure 2 shown.
[0050] The proliferation ability of mesenchymal stem cells in tissue engineered structures after cryopreservation is shown in image 3 shown, the colony-forming ability of mesenchymal stem cells see Figure 4 shown, the three differentiation capabilities of mesenchymal stem cells are shown in Figure 5 shown.
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