# In Vitro Fish Cell Culture: From Primary Muscle Cells to Cell-Based Meat in Cyprinidae

## TL;DR

This review discusses how to grow fish muscle cells in the lab to create sustainable fish meat, focusing on the Cyprinidae family.

## Contribution

The paper provides a comprehensive overview of methods and challenges in cultivating Cyprinidae muscle cells for cell-based meat.

## Key findings

- Common methods for obtaining muscle cells include explant outgrowth and enzymatic dissociation.
- Three-dimensional culture systems and bioreactors show promise for large-scale production.
- Challenges include limited cell lines and high costs of growth media.

## Abstract

Fish provide high-quality protein and important nutrients, but wild fish stocks are declining because of growing food demand and climate change. Cultivated fish meat, which is grown directly from cells, has therefore emerged as a promising approach to support a sustainable and reliable food supply. This review explains the basic structure of fish muscle and describes how scientists grow muscle cells outside the body. We focus particularly on fish from the Cyprinidae family, which includes many widely farmed species. The review outlines common methods for obtaining muscle cells, such as allowing small pieces of tissue to grow out in a dish or using gentle enzymes to separate individual cells. We also describe the conditions needed for these cells to grow, including suitable temperatures, nutrients, and supportive materials that help the cells form muscle-like structures. Recent progress in three-dimensional growing systems and controlled vessels may enable larger-scale production in the future. However, challenges remain, including the limited number of well-characterized Cyprinidae muscle cell types and the high cost of the nutrients required for cell growth. Continued research will help improve these methods and support sustainable food production and aquaculture.

Fish offer an excellent source of high-quality protein with balanced nutrients and low fat content. However, the increasing global demand for food and the impacts of climate change have led to a significant decline in wild fish stocks. Cultivated fish meat has therefore emerged as a promising sustainable food alternative. In this review, we summarize the structural and physiological characteristics of fish muscle and highlight the methods used to establish primary muscle cell cultures, including explant outgrowth and enzymatic dissociation, alongside the optimization of environmental conditions and growth media composition. Particular attention is given to the isolation, development, and characterization of Cyprinidae muscle-derived cell lines via morphological assessments, gene expression profiling, and karyotyping. In addition, we discuss recent advances in scaffold-based and three-dimensional culture systems as well as the application of bioreactors for large-scale production. Current challenges include the limited availability of standardized muscle cell lines, dependence on serum-containing media, and the high cost of growth factors. Future progress will depend on innovations in serum-free formulations, cost-effective media, and reproducible culture protocols. Ultimately, fish muscle cell culture supports not only the development of alternative protein sources but also aquatic health research, disease modeling, and sustainable bioproduction.