| Abstrakt: |
Simple Summary: The adherent and suspension cell culture methods have their own advantages. Adherent culture involves both straightforward steps and inexpensive consumables, and the even spread of cells on the substrate facilitates clear observation of the cell morphology, growth status, and cytopathic effects using a microscope. By comparison, the ability to form a high-density cell population and the ease of scaling up make suspension culture valuable for industrial production, such as vaccine development, bio-pharmaceutical manufacture, medicine screening, and food production. However, very few cell lines can grow in suspension naturally, and the typical commercially used suspension cell culture systems are all mammalian in origin, making them not suitable for a lot of aquatic cases. To meet the needs of aquatic industries, a new ES culture system of aquatic origin was developed through explant outgrowth and enzyme-digesting passaging. The ES cells can adapt to suspension growth naturally and conduct transformation reversibly between the adherent and suspension modes. This culture model was confirmed to be susceptible to different fatal aquatic viruses and suitable for gene engineering as well. As a suspension culture system with no need for microcarriers and special additives, ES cells may be useful for the development of new aquatic serum-free production systems in a bioreactor or become a promising tool for the aquatic vaccine industry. Suspension growth can greatly increase the cell density and yield of cell metabolites. To meet the requirements of aquatic industries, a culture model derived from Anguilla anguilla skin was developed using the explant outgrowth and enzyme-digesting passaging methods. These cells were kept in vitro continuously for over 12 months and subcultured 68 times. This heteroploid cell line, designated as ES, can naturally adapt to adherent and suspension growth reversibly under certain temperatures, serum percentages, and inoculum densities, without the need for any microcarriers or special medium additives. The ES cells can continue being highly productive under a temperature range of 15–37 °C and a serum percentage ranging from 3 to 15%. An inoculum density higher than 5 × 105 cells·mL−1 is necessary for the ES cells to turn into suspension efficiently. The green fluorescent protein (GFP) reporter gene was successfully expressed in the ES cells. The ES cells demonstrated susceptibility to Anguillid herpesvirus (AngHV) and red-spotted grouper nervous necrosis virus (RGNNV). ES is the first natural suspension growth model of aquatic origin; it does not require the processes of suspension domestication and carrier dissolution, making it a promising and cost-effective model for vaccine production, bio-pharmaceutical manufacturing, and cellular agriculture. [ABSTRACT FROM AUTHOR] |
