Cell Line Name
Cell Line Description
The novel SG01 subglottic epithelial cell line has been established to provide a unique resource to investigate subglottic diseases, such as subglottic stenosis. SG01 is an immortalised cell line that can differentiate at the air-liquid interface (ALI) in culture and demonstrates morphology of normal epithelium and cytokeratins of normal epithelium and relevant ion channel expression.
Unique to ECACC
Tissue of Origin
The subglottis is a crucial site for airway infection, inflammation and malignancy. This model provides a unique resource for researchers to study subglottic diseases and potentially test therapeutic agents with a site-specific in vitro model.
Resuscitation: A centrifugation step (100-150g for 2-3 minutes to pellet cells) to remove the cryoprotectant is essential. Rapidly thaw the frozen ampoule in a water bath at 37°C for 1-2 minutes. Transfer the contents to a centrifuge tube and slowly add 5-10ml of pre-warmed growth media. Remove a sample for counting. Centrifuge at 100-150g for 2-3 minutes to pellet cells and seed at 10,000 cells per cm². Subculture Routine: Check cultures daily and media change as recommended. Subculture sub-confluent cultures (maximum 70% confluence) using 0.05% trypsin/EDTA. Incubate for 5-10 min until cells detach. Note: BEGM medium is almost serum-free, therefore use of trypsin inhibitor is essential. Add an equal or greater volume of using growth medium supplemented with 10% FBS to stop digestion. Centrifuge at 200-250g for 5 minutes. The supernatant is removed and the cell pellet re-suspended with fresh BEGM medium prior to transfer to another growth flask. Seed cell into new flasks between 3,000- 10,000 cells per cm². Media will need to be changed more frequently as the culture becomes more established. Cryopreservation: Harvest the cells at 70-80% confluency using trypsin and using growth medium supplemented with 10% FCS as described previously. When the cells are dislodged they are centrifuged at 200g, at 10°C for 7 minutes. The supernatant is discarded and the pellet resuspended in freeze media (45% Conditioned medium, 45% Fresh medium and 10% Dimethyl sulfoxide (DMSO)).
Medium BEGM, also known as LHC-9 with modification (available from Clonetics Corporation, CC3171 (BEBM) plus additives CC4175 or BEGM Bullet Kit, CC3170). Follow manufacturer?s recommendations in preparing the complete growth medium. Note: Additives supplied contain gentamycin which has been omitted for culture at ECACC; media is serum-free.
Country of Origin
Primary subglottic epithelial cells at passage 1 were transfected with a simian virus 40 (SV40) cell immortalization system, utilizing a recombinant lentiviral vector (106 IU/mL) (Applied Biological Materials, Inc., Vancouver, Canada).
Hazard Group (ACDP)
Powell J, Verdon B, Wilson JA, Simpson AJ, Pearson J, Ward C. Establishment of an immortalized human subglottic epithelial cell line. Laryngoscope. 2019 Jan 8. doi: 10.1002/lary.27761. [Epub ahead of print] PubMed PMID: 30623447.
If use of this culture results in a scientific publication, it should be cited in the publication as: SG01 (ECACC 17100230).
Unless specified otherwise, at ECACC we routinely handle all of our cell lines at containment level 2 in accordance with the ACDP guidelines (Advisory Committee on Dangerous Pathogens) (UK). All cell cultures have the potential to carry as yet unidentified adventitious agents. It is the responsibility of the end user to ensure that their facilities comply with biosafety regulations for their own country. ACDP Guidance: Biological agents: Managing the risks in laboratories and healthcare premises.
The Culture Collections represent deposits of cultures from world-wide sources. While every effort is made to ensure details distributed by Culture Collections are accurate, Culture Collections cannot be held responsible for any inaccuracies in the data supplied. References where quoted are mainly attributed to the establishment of the cell culture and not for any specific property of the cell line, therefore further references should be obtained regarding cell culture characteristics. Passage numbers where given act only as a guide and Culture Collections does not guarantee the passage number stated will be the passage number received by the customer.
Cultures supplied by Culture Collections are for research purposes only. Enquiries regarding the commercial use of a cell line are referred to the depositor of the cell line. Some cell lines have additional special release conditions such as the requirement for a material transfer agreement to be completed by the potential recipient prior to the supply of the cell line. Please view the Terms & Conditions of Supply for more information.