Cell & Developmental Biology
Open Access

Advancements in Cell Culture: Optimized Protocol for Crassostrea madrasensis Oyster

Rahem Almoli^{* *}Correspondence: Rahem Almoli, Department of Molecular Microbiology and Immunology, University of South China, Hengyang, China, Email:

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Description

Crassostrea madrasensis, commonly known as the Indian backwater oyster, is a valuable species with significant ecological and economic importance. Research on this species often requires the establishment of cell cultures for various purposes, including physiological studies, biotechnology applications, and conservation efforts. However, developing cell cultures from adult oysters presents unique challenges due to their complex physiological characteristics and the presence of contaminants. In this article, we present an optimized protocol for the routine development of cell culture from adult Crassostrea madrasensis oysters, facilitating research and applications in various fields. Cell culture plays a important role in biological research by providing a controlled environment for studying cellular processes and behaviors outside of the organism. In the case of oysters like Crassostrea madrasensis, cell culture offers opportunities to investigate cellular responses to environmental stressors, infectious diseases, and genetic manipulation. Moreover, cultured cells serve as a valuable resource for producing bioactive compounds, bioremediation, and biotechnology applications.

Challenges in cell culture establishment from adult oysters

Developing cell cultures from adult oysters is inherently challenging due to several factors are

Contamination: Oyster tissues may contain various contaminants, including bacteria, fungi, and protozoa, which can interfere with cell culture establishment and growth.

Low proliferation rates: Adult oyster cells often exhibit low proliferation rates and limited lifespan in vitro, necessary optimization of culture conditions to promote cell growth and viability.

Species-specific requirements: Each oyster species may have unique physiological requirements for cell culture establishment, necessitating species-specific optimization protocols.

Optimized protocol for cell culture development

To address these challenges and facilitate the routine development of cell culture from adult Crassostrea madrasensis oysters, the optimized protocol was

Tissue collection and sterilization:

• Collect oyster tissues (e.g., mantle, gills, adductor muscle) aseptically from healthy adult individuals.

• Rinse the tissues thoroughly with sterile seawater or culture medium to remove debris and surface contaminants.

• Treat the tissues with antimicrobial agents (e.g., antibiotics, antifungal agents) to minimize contamination.

Tissue dissociation and cell isolation:

• Chop the collected tissues into small pieces using sterile scissors or scalpels.

• Digest the tissue fragments using enzymatic solutions (e.g., collagenase, trypsin) to release individual cells from the extracellular matrix.

• Filter the cell suspension through a sterile mesh or sieve to remove undigested tissue fragments.

Culture medium preparation:

• Prepare a culture medium optimized for cell growth, containing essential nutrients, vitamins, and growth factors.

• Supplement the medium with serum or serum substitutes to support cell proliferation and viability.

• Adjust the pH and osmolarity of the medium to physiological levels suitable for oyster cells.

Cell seeding and culture:

• Seed the isolated oyster cells into sterile culture vessels (e.g., tissue culture plates, flasks) at an appropriate density.

• Incubate the cells in a controlled environment with suitable temperature, humidity, and gas exchange.

• Regularly monitor cell growth and morphology under a microscope, and replenish the culture medium as needed.

Subculture and maintenance:

• Subculture the oyster cells periodically to prevent overgrowth and maintain optimal culture conditions.

• Passage the cells at regular intervals using enzymatic or mechanical dissociation methods to promote cell proliferation.

• Perform routine quality control assessments to ensure the absence of contamination and genetic stability of the cell line.

Conclusion

Establishing a reliable and reproducible protocol for the routine development of cell culture from adult Crassostrea madrasensis oysters is essential for advancing research in various disciplines, including aquaculture, environmental science, and biotechnology. By optimizing culture conditions and addressing specific challenges associated with oyster cell culture, researchers can unlock the full potential of Crassostrea madrasensis as a model organism and a valuable resource for scientific inquiry and innovation.