ATCC ATCC Logo 0
  • Careers
  • Support

293SF-3F6

CRL-3537

293SF-3F6 were adapted to suspension serum-free culture. These cells may be used to produce E1-deleted adenoviral vectors as well as used for transfection for recombinant protein production. This product is an ATCC manufactured and accessioned progeny of CRL-12585 cited in US Pat. No. US 6,210,922.
Product category
Human cells
Organism
Homo sapiens, human
Morphology
Lymphocyte-like; single cells to small aggregates
Tissue
kidney
Applications
Bioproduction
Product format
Frozen
Storage conditions
Vapor phase of liquid nitrogen
Buy Now
Price: $844.00 ea
Discounts may be available for our fellow nonprofit organizations. Login to see your price.

Generally ships within 1-3 business days

Documentation

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.

Required Products

These products are vital for the proper use of this item and have been confirmed as effective in supporting functionality. If you use alternative products, the quality and effectiveness of the item may be affected.

Detailed product information

General

Specific applications
Protein production, viral production, adenovirus studies, transfection, recombinant protein production, bioproduction.

Characteristics

Growth properties
Suspension
Derivation
ATCC accessioned progeny of 293G-3F6 cited in US Pat. No. US 6,210,922 as CRL-12585.
Age
embryo
Gender
Female
Comments
293SF-3F6 were adapted from parental 293S cells. Parental 293S cells were adapted to suspension serum-free culture followed by two consecutive cloning by limit dilution. These cells were transformed with adenovirus 5 DNA. 293SF-3F6 cells originate from BRI-NRC, Canada. These cells may be used to produce E1-deleted adenoviral vectors as well as used for transfection for recombinant protein production.

Handling information

Unpacking and storage instructions
  1. Check all containers for leakage or breakage.
  2. Remove the frozen cells from the dry ice packaging and immediately place the cells at a temperature below ­-130°C, preferably in liquid nitrogen vapor, until ready for use.
Complete medium

The base medium for this cell line is HyCell TransFx-H. To make the complete medium add these products to the base medium:

  • 10 mL L-glutamine (ATCC 30-2214) to a final concentration of 4mM
  • 1 mL poloxamer 188 (Fisher Scientific cat # MT13901CI or Corning cat # 13901CI) to a final concentration of 0.2% 
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.

  1. 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).
  2. 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.
  3. Transfer the vial contents to a centrifuge tube containing 9.0 mL complete culture medium and spin at approximately 125 x g for 5 to 7 minutes.
  4. Carefully aspirate the supernatant and discard, leaving the cell pellet.
  5. Gently resuspend the cell pellet in fresh pre-warmed complete growth medium, and transfer cell suspension into a vented, non-baffled shaker flask. Cells should be seeded at a density of 5 x 105 cells/mL.
  6. Place the flask in a 37°C shaking incubator (120 to 130 rpm) with 5-8% CO2.
Subculturing procedure
Re-seed cells at log phase every 2-3 days when the cell density is between 1.5 x 106 and 3.0 x 106 viable cells/mL. Pre-warm fresh growth medium prior to use. Swirl the flask gently to evenly distribute cells in medium. Remove a small volume of cells from the flask and perform cell count. Cultures can be maintained by addition or replacement of fresh medium. Start cultures at 5.0 X 105 cells/mL and maintain between 4 X 105 and 3 X 106 cells/mL.

  1. Seed at 5 x 105cells/mL for a 2 day subculture and 4 x 105 cells/mL for a 3 day subculture (weekend).
  2. To maintain high cell viability, prior to seeding, centrifuge cells for 5min at 170x g.
  3. Discard spent media and re-suspend cell pellet in pre-warmed fresh complete growth media.
  4. Pipette cells gently to break aggregates.
  5. Add the cell suspension to a non-baffled, vented flask.
  6. Place the flask in a 37°C shaking incubator (120 to 130 rpm) with 5-8% CO2.
Medium Renewal: Add fresh medium every 2 to 3 days (depending on cell density). Re-seed cells at log phase every 2-3 days when the cell density is between 1.5 x 106 and 3.0 x 106 viable cells/mL. Cells should be transferred to a new vessel at each media addition. This is because a ring of adhered cells forms on the shaker flask at the liquid air interface. If media were added to the flask above this line, then adhered cells will dislodge and become part of the cell suspension causing reduced viability.
Reagents for cryopreservation
Complete culture medium + 5% DMSO (ATCC 4-X)
Cryopreservation
Complete culture medium + 5% DMSO (ATCC catalog # 4-X). Cells should be frozen at a high concentration (e.g., 7-8 x 106 cells/mL) and at a low passage number. The cells should be ≥ 85% viable prior to freezing.

Quality control specifications

Bacterial and fungal testing
Not detected
Mycoplasma contamination
Not detected
Virus testing
Cytomegalovirus (CMV): Not detected
Epstein-Barr virus (EBV): Not detected
Hepatitis B virus (HBV): Not detected
Human Immunodeficiency virus (HIV): Not detected
Human papillomavirus (HPV): Not detected
Population doubling time
Approximately 24 hrs

History

Deposited as
ATCC accessioned progeny of 293G-3F6 cited in US Pat. No. US 6210922B1 as CRL-12585.

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.

Frequently Asked Questions

References

Need assistance with this product? Contact our Technical Support team.

Telephone

US and Puerto Rico
800-638-6597

Outside the US
+1-703-365-2700

Hours of Operation

Monday-Friday
9:00am - 5:00pm
US Eastern Time