[논문]Principal protocols for the processing of cultured meat

Lee, Seung Yun; Kang, Hea Jin; Lee, Da Young; Kang, Ji Hyeop; Ramani, Sivasubramanian; Park, Sungkwon; Hur, Sun Jin

doi:10.5187/jast.2021.e40

Principal protocols for the processing of cultured meat 원문보기

Journal of animal science and technology : JAST, v.63 no.4, 2021년, pp.673 - 680

Lee, Seung Yun (Department of Animal Science and Technology, Chung-Ang University) , Kang, Hea Jin (Department of Animal Science and Technology, Chung-Ang University) , Lee, Da Young (Department of Animal Science and Technology, Chung-Ang University) , Kang, Ji Hyeop (Department of Animal Science and Technology, Chung-Ang University) , Ramani, Sivasubramanian (Department of Food Science & Biotechnology, Sejong University) , Park, Sungkwon (Department of Food Science & Biotechnology, Sejong University) , Hur, Sun Jin (Department of Animal Science and Technology, Chung-Ang University)

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The purpose of this study was to establish a basic principal procedure for the processing of cultured meat. The first stage involved isolating satellite cells from the desired muscle of an animal using enzymatic digestion (i.e., by using proteases, collagenases, and pronases). The second stage involved culturing the isolated muscle satellite cells in a growth medium containing fetal bovine serum and penicillin/streptomycin with growth factors for an optimal period of time. The second stage involved a basic method for the isolated muscle cells to proliferate while sub-culturing to further induce differentiation in gelatin-coated culture dishes with the general culture medium. The third stage involved the induction of differentiation of muscle satellite cells or formation of myotubes using myogenic medium. Lastly, the fourth stage involved the identification of cell differentiation or myotube formation (myogenesis) using fluorescent dyes. Moreover, the principle of these protocols can be applied to perform primary culture of animal cells. This study will assist beginners with the technical aspects of culturing meat (isolation, cultivation, and differentiation of muscle satellite cells as well as identification of myotube formation for myogenesis).

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표/그림 (4)

그림 Fig. 1. Overview of muscle satellite cell culture.
그림 Fig. 2. Isolation of muscle satellite cells from animals step.
그림 Fig. 3. Muscle satellite cell culture step.
그림 Fig. 4. Diferentiation and identifcation of muscle satellite cells step.

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Analytical grade isoflurane, ethanol (EtOH), bovine serum albumin (BSA), Triton X-100, paraformaldehyde, sucrose, sodium chloride, potassium chloride, sodium phosphate dibasic, potassium phosphate monobasic, 2-methylbutane, protease, and collagenase were purchased from Sigma-Aldrich (St Louis, MO, USA). Dulbeccos’ Modified Eagel’s Medium (DMEM), fetal bovine serum (FBS), horse serum (HS), and penicillin-streptomycin were purchased from Gibco (Thermo Fisher Scientific, Waltham, MA, USA).
Anti-rabbit IgG (H + L) Alexa Fluor 488 conjugate was purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA), and 4´, 6-diamidino-2-phenylindole and dihydrochloride were purchased from Invitrogen (Thermo Fisher Scientific, Waltham, MA, USA).
Briefly, the final medium was prepared with 100 unit/mL of 20% FBS and 100 μg/mL of 1% 1X P/S and stored at 4℃ . Myogenic medium for inducing differentiation was prepared with DMEM- high glucose (HG), HS, and antibiotics. Briefly, the final medium was prepared using 2% HS and 1% 1X P/S in DMEM-HG, and stored at 4℃ until the end of use.
Sterile single-use disposable plastic wares, such as cell strainers (40, 70, and 100 μm), petri dishes, polypropylene centrifuge tubes (15 and 50 mL), and T-flasks (T75), were used

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