The immortalized equine lung cells (extEqFL) represent a significant advancement in veterinary and virological research, primarily for their extended lifespan and utility in studying equine gammaherpesviruses. Developed from primary cells extracted from equine fetal lung by transfection with the retroviral vector LXSN116E6E7, these cells incorporate human papilloma virus oncogenes E6 and E7, enabling propagation beyond the typical 10-12 passages to over 40 passages. This capability allows for prolonged and detailed studies without the rapid senescence associated with primary cells. These cells exhibit comparable transfection efficiency to primary lung cells and maintain stable growth and characteristics throughout extended culture. The extEqFL line is particularly valuable for viral research, offering a consistent and reproducible model for investigating equine herpesvirus infections and their interactions with host cells.
Cell Type
Immortalized Cells
Biosafety Level
2
Depositor
University of Iceland
Disclaimer
1. All test parameters provided in the CoA are conducted using abm's standardized culture system and procedures. The stated values may vary under the end-user's culture conditions. Please verify that the product is suitable for your studies by referencing published papers or ordering RNA (0.5 ?g, Cat.# C207, $450.00) or cell lysate (100 ?g, Cat.# C206, $600.00) to perform preliminary experiments, or alternatively use our Gene Expression Assay Service (Cat# C138). All sales are final. 2. We recommend live cell shipments for ease of cell transfer and this option can be requested at the time of ordering. Please note that the end-user will need to evaluate the feasibility of live cell shipment by taking into account the final destination's temperature variation and its geographical location. In addition, we thoroughly test our cell lines for freeze-thaw recovery. If frozen cells were received and not recovered in your lab under the exact, specified conditions (using recommended culture vessel, media, additional supplements, and atmospheric conditions), a live cell replacement is possible at a cost (plus shipping). 3. All of abm's cell biology products are for research use ONLY and NOT for therapeutic/diagnostic applications. abm is not liable for any repercussions arising from the use of its cell biology product(s) in therapeutic/diagnostic application(s). Please contact a technical service representative for more information. 4. abm makes no warranties or representations as to the accuracy of the information on this site. Citations from literature and provided for informational purposes only. abm does not warrant that such information has been shown to be accurate. 5. abm warrants that cell lines shall be viable upon initiation of culture for a period of thirty (30) days after shipment and that they shall meet the specifications on the applicable abm Material Product Information sheet, certificate of analysis, and/or catalog description. Such thirty (30) day period is referred to herein as the "Warranty Period."
Donor History
5-month old, equine fetus
Expression Region
albumin, AFP, transferrin, LPK, CK8 and CK18
Growth Properties
Adherent, polygonal
Growth Conditions
PriCoat?T25 Flasks (G299) or Applied Cell Extracellular Matrix (G422) are recommended for optimal cell culture.?riGrow III (TM003) + 10% FBS +? mM L-Glutamine (G275) + 600 ?/ml Antibiotic G418 (G271) +?% Penicillin/Streptomycin Solution (G255), 37.0?, 5% CO?.
Quantity
1x106 cells / 1.0 ml
Tissue
Lung
Morphology
Epithelial-like to fibroblast-like
Notes
1. Visually examine the packaging containers for signs of leakage or breakage. 2. Immediately transfer frozen cells from dry ice packaging to a temperature below -130?, preferably in liquid nitrogen vapor phase storage, until ready for use.To ensure the highest level of viability, thaw the vial and initiate culture as soon as possible upon receipt. If continued storage is desired, the vial should only be stored below -130? or in liquid nitrogen vapor phase. Do not store at -70?, as it will result in loss of viability.
Organism
Horse (Equine)
Population Doubling
18 - 28 hours
Species
European Sea Bass (D. labrax)
Propagation
Use of PriCoatTMT25 Flasks (G299) or Applied Cell Extracellular Matrix (G422) is required for cell adhesion to the culture vessels. Grow cells inECM-coated culture vessels unless otherwise specified in the Propagation Requirements below.The base medium for this cell line is Leibovitz's L-15 Medium. To make the complete growth medium, add the following components to the base medium: fetal bovine serum (TM999)* to a final concentration of 10%, Penicillin/Streptomycin Solution (G255) to a final concentration of 1%, L-glutamine (G275) to a final concentration of 2 mM, HEPES to a final concentration of 10 mM, and NaCl to a final concentration of 0.16%.Temperature: 20 - 25
Cryopreservation
Cryopreservation Medium (TM024), or complete growth media with 10% DMSO.
Quality Control
1) Real time PCR; 2) Evaluation of metal cytotoxicity
Applications Range
Research Use Only.
Shipping
Ship with dry ice.
Product Format
Frozen
Product 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.
Storage
Vapor phase of liquid nitrogen, or below -130?.
Thawing Protocol
1. Thaw cells quickly in a 37? water bath while agitating gently (maximum 2 minutes). The vial cap should be kept above the water level to minimize the risk of contamination. 2. Decontaminate the vial by spraying and wiping the exterior of the vial with 70% ethanol. From this point onwards, all operations should be strictly carried out inside a biological safety cabinet using aseptic conditions. 3. Transfer the cell suspension into a 15ml sterile conical tube containing 5ml of pre-warmed, complete growth media. Centrifuge cells at 125xg for 5-7 minutes. 4. Aspirate the supernatant without disturbing the cell pellet. Re-suspend the cell pellet in the recommended pre-warmed, complete growth media and dispense into a T25 culture flask. 5. Incubate the cells at the recommended conditions.
Subculture Protocol
Volumes given below are for a T75 flask; proportionally increase or decrease the volume as required per culture vessel size. Subculture cells once the culture vessel is 80% confluent. 1. Aspirate the culture media, and add 2-3ml of pre-warmed 0.25% Trypsin-EDTA to the culture vessel. 2. Observe the cells under a microscope to confirm detachment (typically within 2-10 minutes). Cells that are difficult to detach can be put in 37?, for several minutes to facilitate detachment. 3. Neutralize Trypsin-EDTA by adding an equal volume of the complete growth media into the culture vessel. 4. Transfer the culture suspension into a sterile centrifuge tube, and centrifuge at 125xg for 5 minutes. The actual centrifuge duration and speed may vary depending on the cell type. 5. Aspirate the supernatant, and re-suspend the pellet with pre-warmed fresh complete growth media. Add appropriate aliquots of the cell suspension to new culture vessels, as desired. 6. Incubate the cells at the recommended conditions.
Reviews of Immortalized Equine Lung Cells (extEqFL)