Three-dimensional scaffold applied to cell culture and preparation method thereof
A three-dimensional scaffold, cell culture technology, applied in the field of cell culture, can solve problems such as speeding up wound repair
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[0044]On the other hand, an embodiment of the present invention provides a method for preparing the three-dimensional scaffold as described above, including:
[0045] Step 1) preparing the acetic acid solution of the collagen-chitosan;
[0046] Step 2) degassing the acetic acid solution of collagen-chitosan and pouring it into a container, freeze-drying, and cutting into a preset shape to obtain a porous scaffold material;
[0047] Alternatively, a cell culture well plate with a plurality of holes is selected, wherein the cell culture well plate satisfies the following conditions: the cell culture well plate includes surrounding wells arranged on the edge and a central well surrounded by the surrounding wells at the center ;
[0048] The acetic acid solution of collagen-chitosan is defoamed and injected into the central hole to a preset height, and water is injected into the surrounding hole to a preset height, and then freeze-dried to obtain a porous scaffold material.
[0...
Embodiment 1
[0080] For convenience of description, the three-dimensional scaffold prepared in Example 1 is designated as A.
[0081] The specific preparation method is as follows:
[0082] 1) prepare the acetic acid solution of collagen-chitosan, wherein, the concentration of collagen is 0.1g / L, and the concentration of chitosan is 1g / L;
[0083] Specifically, first prepare an aqueous solution of acetic acid with a concentration of 0.1M / L, and perform sterile filtration; then mix chitosan with the aqueous solution of acetic acid in a certain ratio, and heat at 45° C. under stirring, Dissolving the chitosan in the aqueous solution of acetic acid to obtain an acetic acid solution of chitosan;
[0084] Then, add the collagen protein solution and water into the acetic acid solution of chitosan in a certain ratio, mix and stir evenly, and obtain the acetic acid solution of collagen-chitosan.
[0085] 2) The obtained collagen-chitosan acetic acid solution was injected into a plate with a diam...
Embodiment 2
[0091] For convenience of description, the three-dimensional scaffold prepared in Example 2 is designated as B.
[0092] The specific preparation method is as follows:
[0093] 1) prepare the acetic acid solution of collagen-chitosan, wherein, the concentration of collagen is 0.1g / L, and the concentration of chitosan is 25g / L;
[0094] Specifically, first prepare an aqueous solution of acetic acid with a concentration of 0.1M / L, and perform sterile filtration; then mix chitosan with the aqueous solution of acetic acid in a certain ratio, and heat at 60° C. under stirring, Dissolving the chitosan in the aqueous solution of acetic acid to obtain an acetic acid solution of chitosan;
[0095] Then, add the collagen protein solution and water into the acetic acid solution of chitosan in a certain ratio, mix and stir evenly, and obtain the acetic acid solution of collagen-chitosan.
[0096] 2) The obtained collagen-chitosan acetic acid solution is injected into 60 central wells of...
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