1. Sale of this item is subjected to the completion of a Material Transfer Agreement (MTA) by the purchasing individual/institution for each order. If you have any questions regarding this, please contact us at [email protected]. 2. 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. 3. 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). 4. 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. 5. 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. 6. 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."
Growth Properties
Adherent, epithelial
Growth Conditions
Use of PriCoat?T25 Flasks (G299) coated with?uman Type IV Placental Collagen is recommended for optimal cell attachment and propagation. Wash with 1X PBS (Mg/Ca) twice times, at least, prior to plating cells.?riGrow X Series Medium (TM5515) + 1% Penicillin/Streptomycin Solution (G255), 37.0?, 5% CO?.Note: warm media to room temperature and protect from light. Media freshness is vital for cell health and propagation.
Quantity
5x105 cells / 1.0 ml
Tissue
Airway
Morphology
Epithelial
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
Human (H. sapiens)
Species
Human
Propagation
Use of PriCoatTM T25 Flasks (G299) or Applied Cell Extracellular Matrix (G422) is required for cell adhesion to the culture vessels. PriGrow X Series Medium (TM5515) + 1% Penicillin/Streptomycin Solution (G255), 37.0°C, 5% CO?.
Cryopreservation
60% complete growth media, 30% FBS and 10% DMSO.
Source Organ
Airway/Lung
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. DO NOT CENTRIFUGE FRESHLY THAWED CELLS. Transfer the vial contents to a 6 well plate with 2 ml fresh, warm complete growth media.?br />4. Incubate the cells at the recommended conditions. 5. Break up cell clumps through gentle pipetting, the next day. Replace media with fresh media, every other day.
Subculture Protocol
Once cells become 75% confluent, detach with Accutase and expand to larger culture vessel, as appropriate. Do not seed cells at very low densities.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 75% confluent. 1. Aspirate the culture media, wash with 5ml 1X PBS without magnesium and calcium. Incubate with 10 ml 1X PBS supplemented with 1 mM EDTA for 3-7 minutes at?7?. 2. Add 5 ml of fresh room temperature Accutase to the culture vessel and let sit at room temperatire for 10 minutes. Periodically observe cells for rounding, using a microscope.?br>3. Detach cells by gently tapping the flask, once the cells appear well-rounded. Collect the cell culture suspension and transfer to 50 ml conical tube without a filter. Pipette the cell solution several times to break up any clumps.?br>4. Apply the cell solution to a filter to strain out clumps' single cell suspension is helpful to achieving a monolayer. Wash cells with 5ml media, and centrifuge at 300xg for 10 minutes.?br>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 Normal Human Primary Small Airway Epithelial Cells