ROM2

CRL-3640 ^™

ROM2 is an adherent epithelial-like cell line derived from a pediatric patient with recurrent otitis media with effusion (inflammatory condition of the middle ear). Due to the characteristic inflammatory gene expression and responses, ROM2 is intended for the study of pediatric recurrent otitis media. Relative to normal non-inflamed pediatric middle ear derived from a patient without otitis media, ROM2 demonstrated greater cytokine expression and response, supporting the retention of an inflamed OM-affected phenotype.

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2 Product Citations

Product category: Human cells
Organism: Homo sapiens, human
Morphology: Epithelial-like
Tissue: Ear; Middle ear; Near eustachian tube orifice
Disease: Otitis media
Product format: Frozen
Storage conditions: Vapor phase of liquid nitrogen

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Documentation

Product sheet

BSL 2

ATCC determines the biosafety level of a material based on our risk assessment as guided by the current edition of Biosafety in Microbiological and Biomedical Laboratories (BMBL), U.S. Department of Health and Human Services. It is your responsibility to understand the hazards associated with the material per your organization’s policies and procedures as well as any other applicable regulations as enforced by your local or national agencies.

ATCC highly recommends that appropriate personal protective equipment is always used when handling vials. For cultures that require storage in liquid nitrogen, it is important to note that some vials may leak when submersed in liquid nitrogen and will slowly fill with liquid nitrogen. Upon thawing, the conversion of the liquid nitrogen back to its gas phase may result in the vial exploding or blowing off its cap with dangerous force creating flying debris. Unless necessary, ATCC recommends that these cultures be stored in the vapor phase of liquid nitrogen rather than submersed in liquid nitrogen.

Detailed Product Information Permits & Restrictions References

General

Specific applications: Pediatric otitis media research

Characteristics

Cells per vial: ≥ 1.0 x 10⁶
Growth properties: Adherent
Age: 5 years
Ethnicity: White
Gender: Male
Clinical data: Derived from a patient with recurrent otitis media with effusion.
Karyotype: 68-81 <3n > ,XXY, + V, + 1, +2, +3,-4, +5,+8, +9,-10,+ 11,add(13)(p11.2),-14, + 15,-16,+ 17, +18,-19, +20,-21, + 1- 3mar[cp 7]
Virus susceptibility: Haemophilus influenzae (Lehmann and Neumann) Winslow et al.
Genes expressed: SV40LT, TNFA, IL1B, IL6, IL8, MUC5AC, MUC5B
Comments: ROM2 is intended for use of study of recurrent otitis media. This cell line was derived from a 5.1 year old white non-hispanic male patient with recurrent otitis media with effusion (inflammatory condition of middle ear). ROM2 was conditionally immortalized by temperature-sensitive mutant tsA58 SV40 Large T antigen; maintenance at 33°C permits expression of SV40 T antigen whereas maintenance at 37°C causes its destruction. This characteristic may permit semi-differentiation under air-liquid interface conditions at 37°C.

Handling information

Temperature

33°C

Temperature effects

Permissive temperature: 33°C

Non-permissive temperature: 37°C

Atmosphere

95% Air, 5% CO₂

Handling procedure

To insure the highest level of viability, thaw the vial and initiate the culture as soon as possible upon receipt. If upon arrival, continued storage of the frozen culture is necessary, it should be stored in liquid nitrogen vapor phase and not at -70°C. Storage at -70°C will result in loss of viability.

Thaw the vial by gentle agitation in a 37°C water bath. To reduce the possibility of contamination, keep the O-ring and cap out of the water. Thawing should be rapid (approximately 2 minutes).
Remove the vial from the water bath as soon as the contents are thawed, and decontaminate by dipping in or spraying with 70% ethanol. All of the operations from this point on should be carried out under strict aseptic conditions.
Transfer the vial contents to a centrifuge tube containing 9.0 mL complete culture medium and spin at approximately 125 to 400 x g for 8 to 12 minutes.
Carefully aspirate the supernatant and discard, leaving the cell pellet.
Gently resuspend the cell pellet in 15 mL fresh pre-warmed complete growth medium, and transfer cell suspension into a vented,collagen I coated T-75 (Corning® Catalog # 356485).
Place the flask in a 33°C incubator with 5% CO₂ .

Subculturing procedure

Volumes used in this protocol are for 75 cm² flask; proportionally reduce or increase amount of dissociation medium for culture vessels of other sizes.

Remove and discard culture medium.
Briefly rinse the cell layer with PBS to remove all traces of serum that contains trypsin inhibitor.
Add 2.0 to 3.0 mL of Gibco™ TrypLE™ Express Enzyme (1X), no phenol red (Thermofisher Scientific catalog # 12604-013) to flask and observe cells under an inverted microscope until the cell layer is dispersed (usually within 5 minutes).
Note: To avoid clumping do not agitate the cells by hitting or shaking the flask while waiting for the cells to detach. Cells that are difficult to detach may be placed at 33°C to facilitate dispersal.
Add 6.0 to 8.0 mL of Trypsin Neutralizing Solution (ATCC PCS-999-004) and aspirate cells by gently pipetting.
Centrifuge the cell suspension at approximately 150 to 400 x g for 8 to 12 minutes to remove dissociation agent.
Resuspend the cell pellet in an appropriate amount of complete culture medium.
Add appropriate aliquots of the cell suspension to new collagen I coated culture vessels.
Cultures can be established between 1.5 x 10⁴ and 4.0 x 10⁴ viable cells/cm².
Incubate cultures at 33°C.

Interval: Maintain cultures at a cell concentration between 1.0 x 10⁴ and 4.0 x 10⁴ cell/cm².

Subcultivation Ratio: A subcultivation ratio of 1:4 is recommended

Medium Renewal: Every 2 to 3 days

Reagents for cryopreservation

Complete culture medium + 10% DMSO (ATCC 4-X)

Quality control specifications

Bacterial and fungal testing: Not detected
Mycoplasma contamination: Not detected
Virus testing: Not detected

History

Depositors: Joseph E. Kerschner, MD
Medical College of Wisconsin
Department of Otolaryngology and Communications Sciences
8701 W. Watertown Plank Rd.
Milwaukee, WI USA 53226
Year of origin: 2023

Legal disclaimers

Intended use

This product is intended for laboratory research use only. It is not intended for any animal or human therapeutic use, any human or animal consumption, or any diagnostic use.

Warranty

The product is provided 'AS IS' and the viability of ATCC^® products is warranted for 30 days from the date of shipment, provided that the customer has stored and handled the product according to the information included on the product information sheet, website, and Certificate of Analysis. For living cultures, ATCC lists the media formulation and reagents that have been found to be effective for the product. While other unspecified media and reagents may also produce satisfactory results, a change in the ATCC and/or depositor-recommended protocols may affect the recovery, growth, and/or function of the product. If an alternative medium formulation or reagent is used, the ATCC warranty for viability is no longer valid. Except as expressly set forth herein, no other warranties of any kind are provided, express or implied, including, but not limited to, any implied warranties of merchantability, fitness for a particular purpose, manufacture according to cGMP standards, typicality, safety, accuracy, and/or noninfringement.

Disclaimers

This product is intended for laboratory research use only. It is not intended for any animal or human therapeutic use, any human or animal consumption, or any diagnostic use. Any proposed commercial use is prohibited without a license from ATCC.

While ATCC uses reasonable efforts to include accurate and up-to-date information on this product sheet, ATCC makes no warranties or representations as to its accuracy. Citations from scientific literature and patents are provided for informational purposes only. ATCC does not warrant that such information has been confirmed to be accurate or complete and the customer bears the sole responsibility of confirming the accuracy and completeness of any such information.

This product is sent on the condition that the customer is responsible for and assumes all risk and responsibility in connection with the receipt, handling, storage, disposal, and use of the ATCC product including without limitation taking all appropriate safety and handling precautions to minimize health or environmental risk. As a condition of receiving the material, the customer agrees that any activity undertaken with the ATCC product and any progeny or modifications will be conducted in compliance with all applicable laws, regulations, and guidelines. This product is provided 'AS IS' with no representations or warranties whatsoever except as expressly set forth herein and in no event shall ATCC, its parents, subsidiaries, directors, officers, agents, employees, assigns, successors, and affiliates be liable for indirect, special, incidental, or consequential damages of any kind in connection with or arising out of the customer's use of the product. While reasonable effort is made to ensure authenticity and reliability of materials on deposit, ATCC is not liable for damages arising from the misidentification or misrepresentation of such materials.

Please see the material transfer agreement (MTA) for further details regarding the use of this product. The MTA is available at www.atcc.org.

Permits & Restrictions

Import Permit for the State of Hawaii

If shipping to the U.S. state of Hawaii, you must provide either an import permit or documentation stating that an import permit is not required. We cannot ship this item until we receive this documentation. Contact the Hawaii Department of Agriculture (HDOA), Plant Industry Division, Plant Quarantine Branch to determine if an import permit is required.

MORE INFORMATION ABOUT PERMITS AND RESTRICTIONS

References

Curated Citations

Blaine-Sauer S, et al. Establishment of novel immortalized middle ear cell lines as models for otitis media. Laryngoscope. 2023 Aug 28;8(5):1428-1435. PubMed 37899851