Blood serum substituting agent and blood serum free rainbow trout embryo cell line culture medium
A serum replacement and embryonic cell technology, applied in embryonic cells, tissue culture, animal cells, etc., can solve the problems of complex purification and identification process of cell products, difficulty in separation and purification of cell products, allergic reactions of heterogeneous serum, etc., to avoid cell Allergic reaction, low price, fast growth effect
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specific Embodiment approach 1
[0010] Embodiment 1: The serum replacement agent in this embodiment consists of 0.3g recombinant human insulin-like growth factor-1 (rhIGF-1), 0.5g recombinant human basic fibroblast growth factor (rh-bFGF), 0.65g platelet-derived Growth factor B (PDGF-B), 1.5g bovine serum albumin (BSA), 5g bovine transferrin, 34g β-mercaptoethanol, 0.294g glutamic acid, 0.110g sodium pyruvate and 2-3g non-essential amino acids.
[0011] The serum replacement agent of this embodiment has a wide range of applications and can be used for serum-free culture of fish and animal cells. The non-essential amino acids in this embodiment were purchased from Shanghai Weike Biochemical Reagent Co., Ltd., which did not contain glutamic acid. The serum replacement agent of this embodiment does not contain cholesterol.
specific Embodiment approach 2
[0012] Specific embodiment two: In this embodiment, the serum-free rainbow trout embryo cell line culture medium is composed of RPMI-1640 medium and the serum replacement agent described in specific embodiment one; every 1000mL serum-free rainbow trout embryo cell line medium contains the specific embodiment. 44.354-45.354 g of the above-mentioned serum replacement agent, and the balance is RPMI-1640 medium.
specific Embodiment approach 3
[0013] Specific embodiment three: present embodiment rainbow trout embryonic cell line culture method:
[0014] 1. Collection of rainbow trout embryos
[0015] In this embodiment, the rainbow trout embryos are obtained from high-quality rainbow trout fertilized eggs of the Bohai Cold Water Fish Experimental Station of the Heilongjiang Fisheries Research Institute of the Chinese Academy of Fishery Sciences.
[0016] 2. Embryo (blastocyst-late gastrulation) cell isolation
[0017] After the collected rainbow trout embryos passed the microscopic examination, they were cultured at 18°C until the late stage of blastula or late gastrula, and then the rainbow trout embryos were repeatedly dipped in 1×PBS solution for 3 to 4 times, and then washed with 70% alcohol Soak for disinfection and sterilization, and then wash three times with 1×PBS solution with a pH value of 7.4, then put it in RPMI-1640 complete medium, use pointed tweezers to peel off the egg membrane to separate the ce...
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