The invention discloses a preparation method and application of cross-linked hydrogel for muscle stem cell culture, and belongs to the technical field of biological food materials. Chitosan, alginate, dextran and Ca<2+> are crosslinked through a physical crosslinking method to form double-network hydrogel with high mechanical strength, and then the hydrogel is coated with heparin and collagen through a dipping coating method, so that the hydrogel can immobilize growth factors and adhere to cells. Meanwhile, the extracted primary muscle stem cells are inoculated onto the hydrogel and cultured for 24 hours in a growth culture medium (79% of DMEM, 10% of FBS and 1% of double antibodies); and then the extracted primary muscle stem cells are cultured in an incubator with a differential medium (97% of DMEM, 2% of horse serum and 1% of double antibodies) for 7 days. By utilizing the hydrogel, the absorption of the muscle stem cells on nutrient substances can be enhanced, and the growth of the muscle stem cells is facilitated. The dual-network hydrogel has the potential to be used as a scaffold for muscle stem cell growth of stem cell culture meat.