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, HeLaH2B-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.

Medium:
Growth Condition:

BioVector NTCC Inc.
www.biovector.net