Cell Cryopreservation

Transfect your cells in bulk, aliquot and cryopreserve to create banks of assay-ready cells without sacrificing performance of the engineered cells in downstream assays.

 

  • Create banks of ‘assay-ready’ cells
  • Overall cell health and transgene expression are not significantly affected
  • Cryopreserve cells immediately or after overnight incubation
  • Can be frozen in a variety of formats including tubes, vials, flasks and multi-well plates

 

 

 

Flexible Timing for Simplified Assay Scheduling

Our scientists have developed several cryopreservation protocols that enable cell archiving while maximizing cell viability and target expression upon thawing. Transfected cells can be cryopreserved following a short 20-minute incubation period or cultured for several hours to overnight prior to cryopreservation. Specific cell types may have slightly increased viability and transfection efficiency using specific cryopreservation conditions, however, overall cell health and transgene expression are not significantly affected. This provides an additional level of flexibility in cell handling post transfection and scheduling of downstream cell applications.

 

 

Insect Cell Cryopreservation

 

 

SH-SY5Y cells were electroporated with 1µg/1E6 cells of pGFP and divided into three groups and incubated for various lengths of time prior to cryopreserving the cells (immediate – 20-minute recovery, brief – 1 hour incubation, O/N – overnight incubation). Cells incubated overnight were harvested via trypsinization prior to freezing. All three cell populations were frozen in a cryopreservation medium containing 90% serum/10%DMSO. Cell viability and transfection efficiency were measured 24 hours post thaw.

Cryopreservation Has No Significant Impact on Downstream Assays

The quality of transfected cells post cryopreservation translates into the maintenance of gene expression and performance in downstream cell-based assays.  Researchers can easily create a bank of assay-ready cells following a single large scale transfection providing an additional level of screen-to-screen reproducibility.

NFκB Activation Assay

 

 

HEK 293T cells were electroporated with an NFκB reporter plasmid and divided into two groups: 1) plated immediately for use in assay, or 2) cryopreserved and assayed post thaw. For the NFκB assays, cells were seeded in 96 well plates (50k cells/well) and treated with TNFα four hours post plating. Cells were incubated overnight and luciferase activity measured.

 

 

Automated Patch Clamp Ion Channel Assay

 

 

8E8 CHO K1 cells were transfected with 1.5µg/1E6 cells of Kv1.5 α-subunit plasmid DNA via flow electroporation.  Transfected cells were cultured at 37°C for 24 hours, transferred to a 28°C incubator for an additional 24 hours, then suspended in freezing medium and cryopreserved in liquid nitrogen using standard methodology.  Thawed cell were assays in single hole (SH) and population patch clamp (PPC) modes on the IonWorks Quattro.  Data courtesy of BioFocus.

 

GPCR Second Messenger Assay

 HEK 293F suspension cells were transfected with a GPCR expression plasmid (1µg/1E6 cells). Half of the transfected cells were cryopreserved 18 hours post transfection. GPCR activity was assayed by cAMP ELISA at 18 hours post electroporation (fresh cells) or at one hour post thawing (frozen cells).

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Cell-based Assays Using the MaxCyte Platform

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