SPINK1 Stable AsPC-1 Cell Line - 1 vial
Name: SPINK1 Stable AsPC-1 Cell Line
Description:
The SPINK1 Stable AsPC-1 Cell Line was stably transduced with lentivirus expressing the pLenti-CMV-SPINK1-2A-GFP plasmid (Cat. No. LVP320652; Accession No. BC025790) to overexpress the human SPINK1 gene and is recommended for studies in pancreatic cancer, and the effects of SPINK1 expression.
Organism: Human (H. sapiens)
Tissue: Pancreas
Donor History: Female, 62, Caucasian, Ascites of a patient with cancer of the pancreas
Growth Properties: Adherent, polygonal
Cell Type: Stable Cell Lines
Storage Condition: Vapor phase of liquid nitrogen, or below -130°C.
Shipping Conditions: Ship with dry ice.
Product Format: Frozen
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.
BioSafety level: II
Growth Conditions: Use of PriCoat™ T25 Flasks (G299) or Applied Cell Extracellular Matrix (G422) is required for cell adhesion to the culture vessels. PriGrow II (TM002) + 10% FBS + 1% Penicillin/Streptomycin Solution (G255), 37.0°C, 5% CO₂. Selection with 2 µg/ml Puromycin (G264)
Unpacking and Storage Instructions:
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°C, 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°C or in liquid nitrogen vapor phase. Do not store at -70°C, as it will result in loss of viability.
Thawing Protocol:
1. Thaw cells quickly in a 37°C 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°C, 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.
Cryopreservation Cryopreservation: Medium (TM024), or complete growth media with 10% DMSO.
Expression Human SPINK1 (detected by Western blot and qPCR)