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Accelerate Cell Line Development

 

The main challenges associated with cell line development include maintaining good cell viability and ensuring monoclonality of isolated cells.

 

You can save till 4 weeks of the overall cloning process!

 

  • Cloning with cellenONE

    Unlike other technologies (microdroplets, limiting dilution, microfluidic traps), with cellenONE, none of the well will contain multiple cells and most will have a single cell present making the process much more efficient.

  • Traditional Cloning

    Traditional cloning that follow Poisson distribution such as limiting dilution are far from efficient. Indeed, several wells within each plate will contain multiple colonies. Due to this, long and tedious multiple cloning steps are required to ensure monoclonality.

BENEFITS

Only single cell dispensed, no doublets

High viability

Avoid recloning

Save time

Save on consumables and media


Unparalleled clonal recovery achieved in 96, 384 and 1536 Microtiter Plates

 

cellenONE as an ideal platform for undertaking automated cloning in both standard (96 MTP) and high density (384 and 1536 MTP) substrates as it ensures monoclonality while maintaining outstanding clonal recovery.

Compatibility with high density substrates offers a dramatic increase in throughput to automatically process the equivalent of 16 individual 96 MTP in under an hour leaving personnel free to perform less routine tasks.

In addition to saving time, one can save on consumables and expensive reagents while ensuring documentation and monoclonality of all isolated cells.

 

Outstanding clonal recoveries were observed with 96% , 94% and 94% of wells containing single colonies in 96, 384 and 1536 MTP, respectively. Additionally, the remaining wells were empty and did not contain multiple colonies.

Monoclonality Verification

 

For most cell line development applications, single cell images recorded in cellenONE’s dispenser just prior to be isolated will be sufficient to prove monoclonality of your cell culture.  

However, if your process requires monoclonality verification post single cell isolation (e.g. immunotherapy development), our team has recently teamed up with scientists at Nexcelom to demonstrate how combining cellenONE® for single cell isolation and Celigo® for automated monoclonality verification could significantly accelerate cell line development.

Both systems uses brightfield and fluorescent cell imaging and are compatible with a wide range of 96, 384, and 1536wp, making these perfect partners for rapid cell line development.
 


Compared to other single cell sorting methods

 

Cost- and time-efficient generation of high-expressing and fast-expanding monoclonal cell lines is critical for the production of biologicals.

Two of the most common methods for cloning cells are:

  • Manual dilution: is highly inefficient and cost consuming as most wells will not contain only a single cell, which forces multiple cloning steps
  • FACS: enables single-cell isolation but suffers from high shear stress and subsequent poor clonal recovery

In order to compare viability and recovery rates between flow cytometry method and cellenONE, single polyclonal mouse hybridoma cells were isolated into 384-well plates.

 

cellenONE® X1 significantly outperforms the FACSAria 3 in all measured aspects of clonal recovery of hybridoma cells

  • Viability

    Gentle piezo acoustic dispensing enables greater outgrowth than discriminating live cells as they are still subject to the high shear stresses of FACS after detection.

  • Recovery

    cellenONE® X1 can process low cell inputs with minimal losses. Total recovery of cells has the potential to be greater than measured as rejected cells are collected into a recovery tube and may be reprocessed.

cellenONE ensures most isolated single cells are maintained alive!


Temperature control improves clonal recovery

 

cellenONE® systems are equipped with a temperature controlled unit to improve clonal recovery.

Interestingly optimal cloning results were obtained at 4ºC for both CHO and HEK cells.

 

CHO Cloning experiment in 384wp at different temperature