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MS40L-low

CRL-3626

MS40L-low is an adherent fibroblast-like cell line orginating from bone marrow stromal cells (MS-5 cells) isolated from a C3H/HeNSIc strain mouse. These cells are transduced and express low levels of Human CD154/CD40L. These cells can function as feeder cells for in vitro culture of human B lymphocytes.
Product category
Animal cells
Organism
Mus musculus, mouse
Classification
Eukaryota, Animalia, Metazoa, Chordata, Vertebrata, Tetrapod
Morphology
Fibroblast-like
Tissue
Bone marrow
Disease
Normal
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
This cell line is suitable for human B lymphocyte research.

Characteristics

Growth properties
Adherent
Age
adult
Strain
C3H/HeNSIc
Antigen expression
CD154 (CD40L)
Comments
MS40L-low has been transduced with Human CD154/CD40L. These cells grow as a monolayer and exhibit potent contact inhibition.

Handling information

Complete medium

The base medium for this cell line is ATCC-formulated IMDM Medium (ATCC 30-2005). 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%
  • GibcoTM 2-Mercaptoethanol (ThermoFisher catalog #21985-023) to a final concentration of 55uM

Note: Because of the limited stability of 2-Mercaptoethanol, we recommend spiking in fresh 2-Mercaptoethanol prior to seeding or performing fluid additions. Media supplemented with 2-Mercaptoethanol should not be stored long term. Complete culture medium is stable for 7 days at 4°C.

 
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 culture medium and spin at approximately 200 to 500 x g for 4 to 8 minutes.
  4. Carefully aspirate the supernatant and discard, leaving the cell pellet.
  5. Gently resuspend the cell pellet with the appropriate amount of growth medium and transfer cell suspension into a vented T-75.
  6. Place the flask in a 37°C incubator with 5% CO2.
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. 
    1. Remove and discard culture medium.
    2. Briefly rinse the cell layer with PBS to remove all traces of serum that contains trypsin inhibitor.
    3. Add 2.0 to 3.0 mL of 0.05% Trypsin-EDTA (Gibco™ 25300-054) to flask and observe cells under an inverted microscope until the cell layer is dispersed (usually up to 30 minutes).
      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 aspirate cells by gently pipetting.
    5. Centrifuge the cell suspension at approximately 200 to 500 x g for 4 to 8 minutes to remove dissociation agent.
    6. Resuspend the cell pellet in an appropriate amount of complete culture medium
    7. Add appropriate aliquots of the cell suspension to new culture vessels.
      Cultures can be established between 1.0 x 104 and 2.0 x 104 viable cells/cm2.
    8. Incubate cultures at 37°C.
  • Interval: Seed cells at subculture at a cell concentration between 5.0 x 103 and 2.0 x 104 cell/cm2.
    Subcultivation Ratio: A subcultivation ratio of 1:3 to 1:5 is recommended.
    Medium Renewal: Every 2 to 3 days
    Reagents for cryopreservation
    Stem Cell Freezing Media (ATCC ACS-3020)

    Quality control specifications

    Bacterial and fungal testing
    Not detected
    Mycoplasma contamination
    Not detected

    History

    Depositors
    Meimei Dong
    California Insititute of Technology
    United States
    Year of origin
    2009

    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

    Curated Citations

    Ohkawara, JI, et al. Culture system for extensive production of CD19+IgM+ cells by human cord blood CD34+ progenitors. Leukemia 12, 764–771 (1998) PubMed: 9593276

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