Cell Seeding Density Calculator

What is Cell Seeding Density?

Cell seeding density refers to the number of cells plated per unit area of the culture surface, typically expressed as cells per square centimeter (cells/cm²). This parameter is critical for reproducible experiments and affects cell behavior, growth kinetics, and experimental outcomes.

Key Formulas

Standard Vessel Areas

Typical Seeding Densities by Cell Type

Different cell types have different optimal seeding densities depending on their size, growth rate, and experimental requirements:

• HeLa cells: 5,000-20,000 cells/cm² (fast-growing cancer cell line)
• HEK293: 20,000-50,000 cells/cm² (loosely adherent, needs higher density)
• CHO cells: 10,000-30,000 cells/cm² (Chinese hamster ovary, industrial)
• Primary cells: 20,000-50,000 cells/cm² (slower growth, limited passages)
• Stem cells: 30,000-100,000 cells/cm² (colony formation, pluripotency)

Why Viability Correction Matters

Cell viability assessment (typically using trypan blue exclusion) ensures you seed the correct number of viable cells. Non-viable cells in your suspension will not attach or proliferate, so the calculator adjusts for viability to ensure your target seeding density reflects only living cells.

For example, if your stock is 1×10⁶ cells/mL with 90% viability, your viable concentration is effectively 9×10⁵ cells/mL. The calculator uses this adjusted value to determine the correct volume to seed.

Best Practices

• Always count cells immediately before seeding
• Use consistent counting methods for reproducibility
• Add 10-15% overage to account for pipetting dead volume
• Mix cell suspensions thoroughly before each aliquot
• Pre-warm media to 37°C before adding to cells
• Document your seeding density and actual plated volumes