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Contributor Information

  • Name Matthew C Holley
  • Institute University of Sheffield
  • Primary citation Lawoko-Kerali et al. 2004. Dev Dyn. 231(4):801-14. PMID: 15499550

Tool Details

  • Tool name: US/VOT-E36 Cell Line
  • Alternate names: US/VOT-E36; Ventral Otocyst-Epithelial cell line number 36; VOT-E36; University of Sheffield/Ventral OTocyst-Epithelial 36
  • Tool type: Cell Lines
  • Tool sub-type: Continuous
  • Organism: Mouse
  • Tissue: Embryonic
  • Disease: Deafness ; Hearing loss
  • Morphology: Epithelial
  • Growth properties: Adherent
  • Model: Transgenic
  • Model description: Conditionally immortalized cell line
  • CRISPR: No
  • Conditional: Yes
  • Conditional description: The cells were derived from ImmortomiceTM (Jat et al 1991 Proc. Nat. Acad. Sci. USA 88, 5096-5100) carrying a stable insertion of the conditional immortalising gene H-2Kb-tsA58, which describes a temperature-sensitive variant of the SV40 immortalising gene that encodes the large tumour antigen under the control of the ?–interferon-sensitive MHC Class 1 promoter.
    The cells proliferating culturing conditions: 33°C with ?IFN; and the cells differentiating culturing conditions: 39°C without ?IFN
  • Application: Inner ear development studies; Gene expression and function of inner ear-specific genes studies; In vitro screening for gene activation and promoter analysis; Ototoxicity (prescribed drugs and agents that ameliorate their affects) studies; Studies on function of inherited deafness mutations; Functional analysis of ion channels, receptors and signalling pathways in vitro
  • Description: US/VOT-E36 cells represent epithelial progenitors with potential to differentiate into sensory and nonsensory epithelial cells.
    The conditionally immortal cell line was established from the ventral otocyst of the Immortomouse at embryonic day 10.5 (plug in mouse designated E0.5 and birth at E18-19). At this stage the sensory epithelia have not differentiated, and the epithelium is competent to form most of the cells within the cochlear duct, including primary sensory neurons. VOT-E36 cell line has been characterised extensively by timed expression of epithelial cells and inner ear sensory cells markers in conjunction with the transcription factor GATA3 under differentiating conditions in vitro.
    It has also been screened with Affymetrix mouse Micro-arrays. It forms cells of various cochlear phenotypes under differentiating conditions in vitro and despite being derived from an earlier developmental stage, it produces more advanced states of sensory cell differentiation than occurs with UB/OC-1, UB/OC-2
  • Research area: Developmental biology; Drug development; Genetics
  • Production details: Homozygous male Immortomice (originally derived from injected oocytes of CBA/Ca x C57BL/10 mice) were time-mated with wild-type C57Bl/6 female mice to produce heterozygous offspring. Animals were killed by cervical dislocation, in accordance with UK Home Office regulations. Otocysts were removed from E10 embryos under sterile conditions and then dissected to isolate the ventral region. Further selection of ventral otocyst explants was based on expression of the immortalizing gene (conditions: medium containing 50 U/ml ?-interferon (?IFN) at 33°C) and tissue-specific markers in conjunction with the transcription factor GATA3 under differentiating conditions in vitro
  • Cellosaurus ID: CVCL_VC36

  • For Research Use Only

Target Details

Application Details

  • Application: Inner ear development studies; Gene expression and function of inner ear-specific genes studies; In vitro screening for gene activation and promoter analysis; Ototoxicity (prescribed drugs and agents that ameliorate their affects) studies; Studies on function of inherited deafness mutations; Functional analysis of ion channels, receptors and signalling pathways in vitro

Handling

  • Format: Frozen
  • Growth medium: MEM with 10% FCS, 50 Units/ml y-IFN, L-glutamine
  • Temperature: 33 C
  • Volume: 1 ml
  • Storage medium: Cytiva HyCloneTM FetalClone™ II Serum (U.S.) Thermo Fisher
  • Storage conditions: Liquid Nitrogen
  • Shipping conditions: Dry ice
  • Characterisation tests:
    Affymetrix mouse Micro-arrays;
    Proliferation (dependent upon the T-antigen);
    Morphology (at differentiating conditions the cells formed a uniform, confluent monolayer with a squamous epithelial morphology);
    Tissue type (GATA3, Cytokeratin, ?4 integrins; ?3-tubulin, Pax-2, Myosin VI and VIIa expression tests)
  • Mycoplasma free: Yes
  • Biosafety level: 1

Documentation

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References

  • •  Holley et al. 2007. Hear Res. 227(1-2):32-40. PMID: 16797894
  • •  Helyer et al. 2007. Eur J Neurosci. 25(4):957-73. PMID: 17331193
  • •  Liu et al. 2006. Otol Neurotol. 27(3):414-21. PMID: 16639283
  • •  Lawoko-Kerali et al. 2004. Dev Dyn. 231(4):801-14. PMID: 15499550