The invention provides a preparation method of a
cell culture three-dimensional
scaffold and
cell culture meat, which comprises the following steps: firstly, preparing a
gelatin-
carrageenan mixed solution and a gel precursor solution of transglutaminase, dividing into two parts, incubating at constant temperature to obtain two parts of composite hydrogel, randomly freeze-
drying one part to obtain
aerogel, and freeze-
drying the
aerogel to obtain the
cell culture three-dimensional
scaffold. Stacking on the other part and incubating at constant temperature again to obtain the prefabricated double-layer gel. The prefabricated double-layer gel is directionally frozen,
ice crystals grow upwards from the bottom, freezing is stopped after the temperature is reduced to the set temperature, and the
cell culture three-dimensional
scaffold is obtained through
drying. The
stent adopts an ordered-disordered partition design, parallel micro-channels are formed in the lower layer by using a directional ice template method, and
muscle fiber arrangement is simulated; a porous spongy structure is constructed on the upper layer by using a random ice template method, and is adapted to
adipocyte proliferation. Two
layers of interfaces are interpenetrated and communicated, natural mechanical gradients are combined, natural tissue characteristics are re-engraved, the traditional limitation is broken through, the cell co-culture efficiency and the
cultured meat quality are improved, and industrialization is promoted.