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BECK-1

CRL-3384

Product category
Human cells
Organism
Homo sapiens
Morphology
Rounded
Tissue
Bone; Marrow
Disease
Lymphoblastic Leukemia
Product format
Frozen
Storage conditions
Vapor phase of liquid nitrogen
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Documentation

Biosafety Icon BSL 2

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.

RPMI-1640 Medium

30-2001

Price: $22.00 ea

Fetal Bovine Serum (FBS)

30-2020

Price: $645.00 ea

L-Glutamine Solution, 200 mM

30-2214

Price: $19.00 ea

Dimethylsulfoxide (DMSO)

4-X

Price: $49.00 ea

Detailed product information

General

Specific applications
Useful for studying hypodiploid ALL, Bcl-2 member signaling, and near-haploid cytogenetics.

Characteristics

Cells per vial
Approximately 7.0 x 106 to 1.0 x 107
Volume
1.0 mL
Growth properties
Suspension
Gender
Female
Tumorigenic
NRAS G12S, CDKN2A/B d
Comments

Cells possess an NRAS G12S mutation and a CDKN2A/B deletion.

Cells lack expression of Bim. Immunophenotyping done by Cy-TOF.

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 RPMI-1640 (ATCC 30-2001). To make the complete medium add the following components: 

  • Fetal bovine serum (FBS; ATCC 30-2020) to a final concentration of 20%  
  • L-Glutamine (ATCC 30-2214) to a final concentration of 4 mM
  • HEPES (Thermo Fisher cat# 15630-080) to a final concentration of 25 mM
Temperature
37°C
Atmosphere
95% Air, 5% CO2
Handling procedure

To ensure 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. Resuspend cell pellet with the recommended complete medium (see the specific batch information for the culture recommended dilution ratio). It is important to avoid excessive alkalinity of the medium during recovery of the cells.  It is suggested that, prior to the addition of the vial contents, the culture vessel containing the complete growth medium be placed into the incubator for at least 15 minutes to allow the medium to reach its normal pH (7.0 to 7.6). pH (7.0 to 7.6).
  5. 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.

         

Handling notes

CRL-3384 cells will sink to the bottom of the vessel over time.

Roller bottles and shaker flasks have not been tested on these cells; however, the constant agitation should prevent the cells from sinking and becoming too dense, as they can in stationary. Stacked vessels and/or roller bottles should not be used.

If seeding at the low end of the range AND using the minimum vessel volume, the vessel should be stood on the short end to prevent a drop in viability.

Subculturing procedure
Volumes used in this protocol are for 75 cm2 flask; proportionally reduce or increase amount of dissociation medium for culture vessels of other sizes. Corning® T-75 flasks (catalog #430641) are recommended for subculturing this product.
  1. Culture to be grown via shaker flasks with incubator humidity. Shaking RPM: 130 .
  2. Observe cells under an inverted microscopes).
  3. Use Corning Recommended Shaker Flask Volumes
  4. Add appropriate aliquots of the cell suspension to new culture vessels.
    Cultures can be established between 8.0 x 105 and 1.0 x 106 viable cells/cm2.
  5. Incubate cultures at 37°C.
Interval: Maintain cultures at a cell concentration between 2.0 X 105 and 3.0 X 106 cell/cm2.
Subcultivation Ratio: A subcultivation ratio of 1:3 to 1:8 is recommended
Medium Renewal: 2 to 3 times per week
Reagents for cryopreservation
81.5% RPMI-1640 + 10% FBS + 2% L-Glutamine + 1.5% HEPES + 5% DMSO

Quality control specifications

Bacterial and fungal testing
Not detected
Mycoplasma contamination
Not detected
Virus testing
Hepatitis B virus (HBV): Not detected
Cytomegalovirus (CMV): Not detected
Human Immunodeficiency virus (HIV): Not detected
Epstein-Barr virus (EBV): Not detected
Human papillomavirus (HPV): Not detected
STR profiling
Amelogenin: X
CSF1PO: 11
D13S317: 12
D16S539: 12
D5S818: 12
D7S820: 12
THO1: 9
TPOX: 11
vWA: 17

History

Depositors
E Diaz-Flores
Year of origin
2014

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.

References

Curated Citations

Diaz-Flores E, etal. BCL-2 as a therapeutic target for hypodiploid B-lineage acute lymphoblastic leukemia". Cancer Res 79(9):2339-2351, 2019. PubMed: 30862722

Session 614. Acute Lymphoblastic Leukemia: Therapy, excluding Transplantation: Chemo Immuno and Targets--Improving Combinations in ALL" ASH Annual Meeting - Invited Program Plus Oral Abstracts, 2019. 

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