Differences Between Primary Cell Culture and Subculture

News 8 5 月, 2025

Primary culture refers to directly obtaining tissue or organ samples from an organism and culturing them; it’s also called initial culture. The culture is maintained outside the organism without splitting or transferring during the process.
Subculture (passage culture) involves splitting cultured cells into smaller portions and transferring them into new vessels for continued growth.

In primary culture, animal tissues are removed, dissociated into single cells (commonly using trypsin), and placed into suitable culture media to survive, grow, and proliferate. The process is relatively complex with strict sterile conditions required throughout.
In subculture, cells from primary culture are further digested (e.g., with trypsin) and transferred to continue culturing. Subcultured cells retain the same karyotype as primary cells but can divide indefinitely in vitro. They generally survive easily under standard conditions and proliferate well but still require sterile technique and precautions to avoid cross-contamination.

Primary cells will divide but typically only up to 10 passages before growth slows, and most cells become senescent or die. Cells attach to the culture surface within hours and can form a monolayer in 5–7 days if seeded at the right density.
Subcultured cells can typically be passaged 40–50 times. They attach within 2 hours and can form a monolayer in 2–4 days, after which they require further passaging.

Primary culture provides a powerful tool for studying cellular growth, metabolism, and proliferation, while also laying the groundwork for subcultures. It’s used in clinical applications, such as culturing tumor cells from surgical specimens to screen anti-cancer drugs and guide chemotherapy selection.
Subculture is widely used in medical fields, including reconstructing complex structures (e.g., tooth roots in dentistry) and improving amniotic cell culture techniques. Subculture allows multiple rounds of cloning, increasing success rates and providing more cells for karyotype analysis, enhancing prenatal diagnostics.