Sf9 Cell Lines
Succeed in Developing Monoclonal Sf9 Cell Lines
The baculovirus expression vector system (BEVS), one of the most versatile eukaryotic vector systems, enables the in vitro expression of heterologous genes in insect cells. However, generating insect Sf9 clones for recombinant protein production has been almost impossible due to unique challenges presented by Sf9 cells with traditional methods, such as limiting dilutionA statistical cloning approach where cells are diluted to low density to “hopefully” seed one cell per well; commonly used but can be low-efficiency for difficult-to-clone lines. More and droplet-based sorting.
Traditional Sf9 cell line development
- High Costs – Limiting dilutionA statistical cloning approach where cells are diluted to low density to “hopefully” seed one cell per well; commonly used but can be low-efficiency for difficult-to-clone lines. More and sorting are expensive and labor-intensive
- Low To No Viability – Stressors, such as low seeding density and high-pressure sorting reduce cell survival of highly sensitive cells
- Unwanted Protein Production Variability – Expansion using polyclonal cells creates unstable cell lines with unreliable protein production
- Increased Efficiency – Huge reduction in consumable usage and less than 2 hours of hands-on time
- Clones That Grow – Gentle isolation by gravity combined with shared culture media exponentially increase survival and growth
- MonoclonalityEvidence that a colony/line originated from one cell; often supported by time-course imaging and traceability. More – Expansion of truly monoclonal colonies into stable cell lines
Sf9 clonal outgrowth efficiency from isolated CellRafts and limiting dilution.
Learn how to quickly and affordably generate monoclonal Sf9 production cell lines
Scientific Poster
A scientific poster comparing the development of cell lines from Sf9 cells using an optimized CellRaft AIR workflow compared to limiting dilutionA statistical cloning approach where cells are diluted to low density to “hopefully” seed one cell per well; commonly used but can be low-efficiency for difficult-to-clone lines. More.
Application Note
An application note detailing the CellRaft AIR protocol for the development and expansion of monoclonal cell lines from Sf9 cells.
Software-guided Identification and Automated Isolation of Cells of Interest
CellRaftA microscale polystyrene growth surface within an array used to spatially segregate cells/colonies while maintaining shared media access, enabling imaging over time and targeted isolation. More Cytometry™ software allows for image-based verification of single cells to ensure monoclonalityEvidence that a colony/line originated from one cell; often supported by time-course imaging and traceability. More and analysis of a variety of parameters over time, ranging from size to morphology to gene expression. Users can easily define the characteristics of the target cells or colonies and map them for software-guided CellRaftA microscale polystyrene growth surface within an array used to spatially segregate cells/colonies while maintaining shared media access, enabling imaging over time and targeted isolation. More selection and automated isolation by the CellRaft AIR SystemAn all-in-one platform for imaging, identifying, and isolating viable single cells, colonies, or organoids using CellRaft Arrays and software-guided selection with CellRaft Cytometry. More.
Image Cells
[glossary]Cells are plated on a CellRaft® Array using standard plating techniques, eliminating the need for trypsin, scraping, high-pressure fluidics, and limiting dilution. The array is scanned, imaged, and recorded, allowing the user to track cells as they grow over time. [/glossary]
Grow Cells
[glossary]Target cells grow unperturbed in optimized conditions in the imaging platform, increasing cell viability, and downstream read depth. The CellRaft Arrays allow different media, compound dosing, and growth conditions to be tested on a single consumablepropriety culture dish. [/glossary]
Isolate Cells
[glossary]When target cells are ready for collection, the gentle release mechanism leaves cells unperturbed, so transfer to a PCR or culture plate won’t affect the quality of the cells for downstream analysis. Cells are also moved into lysis buffer quickly, preventing the degradation of genetic material in sensitive cell lines.[/glossary]
