Description: HeLa cells dually transduced with Retrovirus to stably express H2B tagged with GFP, from pWZL backbone, and H2B tagged with mCherry, both fluorophores sorted for low expression.

Description Key Words: HeLa H2B tagged GFP mCherry

Also Known As:   HeLa-2FP HeLa^H2B-2FP“, HeLa_H2B-GFP_H2B-mCH, HeLa–H2B GFP and H2B mCH  

Organism: Human (Homo sapiens) 

Strain: N/A

Tissue: Cervix cervical carcinoma

Growth Properties: Adherent

Morphology: Epithelial like

Growth Medium:

DMEM – high glucose, 10% BGS, 1% NEAA, 1% Glutamax

100ug/mL Hygromycin (H2B-GFP) and 300ug/mL G418 (H2B-mCh)

Resuscitation: Remove protective cryoflex layer around the ampoule prior to thawing. Thaw the ampoule by gently agitating in a 37°C waterbath; thawing should be rapid (around 2 minutes). A centrifugation step to remove the cryoprotectant after thawing is necessary for this cell line. Recovery from thaw may take 2 days.

Subculturing Procedure:

Medium Renewal: 2-3 times per week. 

Subcultivation Ratio: 1:8 – 1:16 Seeding density 0.8 – 1.0 x10⁴cells/cm². Split subconfluent cultures (70-80%). Harvest the cells using 0.05% Trypsin/EDTA at 37°C for 5 minutes.

Culture Conditions: Incubate the culture at 37°C with 10% CO₂.

Cryoprotectant Medium: 10% DMSO + 90% FCS

Handling Procedure for Frozen Cells: Upon receipt, frozen ampoules should be transferred directly to liquid nitrogen storage without delay, if not to be used immediately. Storage at -80°C may result in loss of viability.

Additional Information: One of a pair of HeLa derivatives genetically modified to carry fluorescently tagged histone H2B proteins. H2B is a core component of the nucleosome, where it plays an important role in transcriptional regulation, DNA repair, and other essential cellular functions. One cell line, HeLa H2B-GFP, carries H2B tagged with eGFP. This cell line, HeLa-2FP, carries both a H2B tagged with eGFP and H2B tagged with mCherry. Expression of both proteins enabled study of chromatin compaction in live cells using FLIM-FRET microscopy. The cell lines were published in PNAS in 2019, where they were used to study the remodelling of chromatin architecture at double-strand break sites during DNA repair.

Depositor: Tony Cesare, Children's Medical Research Institute, Westmead Australia

Reference: Lou J et al. Phasor histone FLIM-FRET microscopy quantifies spatiotemporal rearrangement of chromatin architecture during the DNA damage response. Proc Natl Acad Sci U S A (2019) 116(15): 7323-7332. doi: 10.1073/pnas.1814965116.

PMID: 30918123

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