Crustacean cell culture: historical developments, methodological advances, and future directions

Area of Science:

• Marine invertebrate biology
• Cell culture technology
• Invertebrate immunology

Background:

• Crustaceans are ecologically vital marine invertebrates, but their in vitro research is limited by the lack of immortal cell lines.
• Historical challenges include slow proliferation, complex nutritional needs, osmotic sensitivity, and different cell cycle regulation compared to vertebrates.
• Hemocytes are the most studied crustacean cells, but their heterogeneity and sensitivity hinder long-term in vitro viability.

Purpose of the Study:

• To review historical developments and recent advances in crustacean cell culture.
• To identify critical technological gaps for establishing sustainable crustacean cell lines.
• To highlight progress in creating stable and functional in vitro models for crustaceans.

Main Methods:

• Integration of stem cell-oriented approaches.
• Optimization of tissue-specific media and osmolarity.
• Application of transcriptomic and single-cell sequencing for molecular characterization.
• Exploration of three-dimensional culture systems and extracellular matrix scaffolds.

Main Results:

• Improved longevity, phenotypic stability, and functional maintenance of crustacean cell cultures.
• Identification of hematopoietic progenitor-type and stem-like cells.
• Enhanced cell attachment and differentiation potential using novel culture systems.

Conclusions:

• Recent methodological progress has significantly advanced crustacean primary cell culture.
• Addressing technological gaps is crucial for developing reproducible and sustainable crustacean cell lines.
• These advancements hold promise for expanding in vitro research applications for crustaceans.