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BJ

CRL-2522

BJ cells are fibroblasts established from skin taken from normal foreskin from a neonatal male. Applications include toxicology research.
Product category
Human cells
Organism
Homo sapiens, human
Cell type
fibroblast
Morphology
fibroblast
Tissue
Skin; Foreskin
Disease
Normal
Applications
3D cell culture
High-throughput screening
Toxicology
Product format
Frozen
Storage conditions
Vapor phase of liquid nitrogen
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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
The cells may be used for stable transfection studies.

Characteristics

Growth properties
Adherent
Passage history
A frozen ampule at population doubling 2.3 was received at the ATCC in April, 2000.
Derivation
The line was established from skin taken from normal foreskin.
Age
neonate
Gender
Male
Comments
The BJ cell line has a long lifespan in comparison with other normal human fibroblast cell lines.
A frozen ampule at population doubling 2.3 was received at the ATCC in April, 2000.
These cells have a reported normal diploid karyotype at population doubling 61 but an abnormal karyotype at population doubling 82.
They are telomerase negative.
Cells from ATCC ampules have the capacity to proliferate to a maximum of 72 population doublings before the onset of senescence.

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 ATCC-formulated Eagle's Minimum Essential Medium (ATCC 30-2003). To make the complete growth medium, add the following components to the base medium: fetal bovine serum (ATCC 30-2020) to a final concentration of 10%.
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. Unscrew the top of the vial and transfer the contents to a sterile centrifuge tube containing 9 mL of the recommended medium. Remove the cryoprotectant agent (DMSO) by gentle centrifugation 150 to 400 x g for 8 to 12 minutes. Discard the supernatant, and resuspend the cells in 1 or 2 mL of complete growth medium. Transfer the cell suspension into the culture vessel containing the complete growth medium and mix thoroughly by gentle rocking. It is recommended that the cryoprotective agent be removed immediately.  Resuspend the cell pellet in 8-10 ml of the recommended complete medium and dispense into a 25 cm2 culture flask. Mix thoroughly by gentle rocking. 
  4. Incubate the culture at 37°C in a suitable incubator.  A 5% CO2 in air atmosphere is recommended if using the medium described on this product sheet.

 

Subculturing procedure
Volumes are given for a 75 cm2 flask. Increase or decrease the amount of dissociation medium needed proportionally for culture vessels of other sizes. Corning® T-75 flasks (catalog #430641) are recommended for subculturing this product.
  1. Remove and discard culture medium.
  2. Briefly rinse the cell layer with D-PBS (ATCC 30-2200) to remove all traces of serum that contains trypsin inhibitor.
  3. Add 2.0 to 3.0 mL of Trypsin-EDTA solution (ATCC 30-2101) to flask and observe cells under an inverted microscope until cell layer is dispersed (usually within 5 to 15 minutes).
    Note: To avoid clumping do not agitate the cells by hitting or shaking the flask while waiting for the cells to detach. Cells that are difficult to detach may be placed at 37°C to facilitate dispersal.
  4. Add 6.0 to 8.0 mL of complete growth medium and aspirate cells by gently pipetting.
  5. Add appropriate aliquots of the cell suspension to new culture vessels.
  6. Incubate cultures at 37°C.
Subcultivation Ratio: A subcultivation ratio of 1:2 to 1:9 is recommended
Medium Renewal: Every 2 to 3 days
Reagents for cryopreservation
Complete growth medium supplemented with 5% (v/v) DMSO (ATCC 4-X)

Quality control specifications

Bacterial and fungal testing
Not detected
Mycoplasma contamination
Not detected
Population doubling capacity
≥ 70 PDL
STR profiling
Amelogenin: X,Y
CSF1PO: 10,12
D13S317: 8,9
D16S539: 9,13
D5S818: 12
D7S820: 11,12
TH01: 7,8
TPOX: 10,11
vWA: 16,18
D3S1358: 14,16
D21S11: 29
D18S51: 17,19
Penta_E: 7,17
Penta_D: 12,13
D8S1179: 9,11
FGA: 22,23
D19S433: 14
D2S1338: 16,20

History

Deposited as
human
Depositors
JR Smith

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.

Permits & Restrictions

Import Permit for the State of Hawaii

If shipping to the U.S. state of Hawaii, you must provide either an import permit or documentation stating that an import permit is not required. We cannot ship this item until we receive this documentation. Contact the Hawaii Department of Agriculture (HDOA), Plant Industry Division, Plant Quarantine Branch to determine if an import permit is required.

MORE INFORMATION ABOUT PERMITS AND RESTRICTIONS

Frequently Asked Questions

References

Curated Citations

Yi X, et al. Both transcriptional and posttranscriptional mechanisms regulate human telomerase template RNA levels. Mol. Cell. Biol. 19(6): 3989-3997, 1999. PubMed: 10330139

Bodnar AG, et al. Extension of life-span by introduction of telomerase into normal human cells. Science 279: 349-352, 1998. PubMed: 9454332

Huffman KE, et al. Telomere shortening is proportional to the size of the G-rich telomeric 3'-overhang. J. Biol. Chem. 275: 19719-19722, 2000. PubMed: 10787419

Morales CP, et al. Absence of cancer-associated changes in human fibroblasts immortalized with telomerase. Nat. Genet. 21: 115-118, 1999. PubMed: 9916803

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