KC-6517

CT26-CD20-GFP-Luc2-Medium Cell Line

64118

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Background of CT26-CD20-GFP-Luc2-Medium Cell Line

CD20 encodes a member of the membrane-spanning 4A gene family. Members of this nascent protein family are characterized by common structural features and similar intron/exon splice boundaries and display unique expression patterns among hematopoietic cells and nonlymphoid tissues. This gene encodes a B-lymphocyte surface molecule which plays a role in the development and differentiation of B-cells into plasma cells. This family member is localized to 11q12, among a cluster of family members. Alternative splicing of this gene results in two transcript variants which encode the same protein.

Specifications

Catalog Number	KC-6517
Cell Line Name	CT26-CD20-GFP-Luc2-Medium Cell Line
NCBI/UniProt Accession Number	NM_152866.3
Clone Number	9#
Host Cell Line	CT26
Description	CT26 cell line stably expressing exogenous CD20-GFP-Luc2 gene
Quantity	One vial of frozen cells (≥2-10⁶/vial)
Stability	Stable in culture over a minimum of 10 passages
Application	Drug screening and biological assays
Freezing Medium	70% RPMI1640 + 20% FBS + 10% DMSO
Propagation Medium	RPMI1640 + 10% FBS
Selection Marker	NA
Morphology	Epithelial
Subculture	Split saturated culture 1:4-1:8 every 2-3 days; seed out at about 1-2 × 10⁵ cells/mL
Incubation	37 °C with 5% CO₂
Storage	Liquid nitrogen immediately upon receiving
Doubling Time	Approximately 30 hours
Mycoplasma Status	Negative

Cell Line Generation

CT26-CD20-GFP-Luc2-Medium cell line was generated using a lentiviral vector expressing the CD20-GFP-Luc2 sequence.

Characterization

Figure 1: Characterization of CD20 overexpression in CT26-CD20-GFP-Luc2-Medium stable clones using FACS.

Figure 2: Characterization of GFP overexpression in CT26-CD20-GFP-Luc2-Medium stable clones using FACS.

Figure 3: Characterization of the CT26-CD20-GFP-Luc2-Medium Cell Line stable clone using Bright-Lite Luciferase Assay System in the conditions of different cell number.

Figure 4: Characterization of CT26-CD20-GFP-Luc2-Medium cell line stable clone using PCR sequencing.

Cell Resuscitation

Prewarm culture medium (RPMI1640 supplemented with 10% FBS) in a 37°C water bath.
Thaw the frozen vial in a 37°C water bath for 1-2 minutes.
Transfer the vial into biosafety cabinet, and wipe the surface with 70% ethanol.
Unscrew the top of the vial and transfer the cell suspension gently into a sterile centrifuge tube containing 9.0 mL complete culture medium.
Spin at ~ 125 × g for 5-7 minutes at room temperature, and discard the supernatant without disturbing the pellet.
Resuspend cell pellet with the appropriate volume of complete medium and transfer the cell suspension into a T25 culture flask.
Incubate the flask at 37°C, 5% CO₂ incubator.
Split saturated culture 1:4-1:8 every 2-3 days; seed out at about 1-2 × 10⁵ cells/mL.

Cell Freezing

Prepare the freezing medium (70% RPMI1640 + 20% FBS + 10% DMSO) fresh immediately before use.
Keep the freezing medium on ice and label cryovials.
Transfer cells to a sterile, conical centrifuge tube, and count the cells.
Centrifuge the cells at 250×g for 5 minutes at room temperature and carefully aspirate off the medium.
Resuspend the cells at a density of at least 3×10⁶ cells/mL in chilled freezing medium.
Aliquot 1 mL of the cell suspension into each cryovial.
Freeze cells in the CoolCell freezing container overnight in a -80°C freezer.
Transfer vials to liquid nitrogen for long-term storage.

References

Abrahamsen IW, Kjellevoll S, Greve-Isdahl M, Mensali N, Wälchli S, Kumari S, Loland BF, Egeland T, Kolstad A, Olweus J. T cells raised against allogeneic HLA-A2/CD20 kill primary follicular lymphoma and acute lymphoblastic leukemia cells. Int J Cancer. 2012 Apr 15;130(8):1821-32. doi: 10.1002/ijc.26209. Epub 2011 Aug 12. PMID: 21630262.

Use License Agreement

Research Use Only.

Not for use in diagnostic procedures or therapeutic applications.

Redistribution of the cell line or its derivatives is prohibited without prior written permission from Kyinno Biotechnology.