D2N

TIB-58 ^™

Explore our data

Product category: Animal cells
Organism: Mus musculus, mouse
Classification: Eukaryota, Animalia, Metazoa, Chordata, Vertebrata, Tetrapod
Morphology: lymphoblast
Tissue: Spleen
Disease: Leukemia
Applications: 3D cell culture

Immune system disorder research
Product format: Frozen

See Additional Product Information

Price: $803.00 EA

Discounts may be available for our fellow nonprofit organizations. Login to see your price.

Generally ships within 1-3 business days

Documentation

Product sheet

BSL 1

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.

General

Specific applications: This line was derived from a leukemia induced by N-tropic Friend murine leukemia virus.
The cells are negative for surface immunoglobulin (sIg-) and Thy-1.2 (Thy-1.2-).
Tested and found negative for ectromelia virus (mousepox).

Characteristics

Growth properties: Suspension
Derivation: This line was derived from a leukemia induced by N-tropic Friend murine leukemia virus.
Strain: DBA/2
Tumorigenic: Yes;
Yes, in DBA/2 mice
Antigen expression: H-2d; Ia8
Genes expressed: H-2d; Ia8
Comments: This line was derived from a leukemia induced by N-tropic Friend murine leukemia virus.
The cells are negative for surface immunoglobulin (sIg-) and Thy-1.2 (Thy-1.2-).
Erythroid differentiation and hemoglobin synthesis are NOT inducible by DMSO.
Tested and found negative for ectromelia virus (mousepox).

Handling information

Unpacking and storage instructions

Check all containers for leakage or breakage.
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

RPMI 1640 medium with 0.01 mM 2-mercaptoethanol, 90%; heat- inactivated fetal bovine serum, 10%

Handling procedure

HANDLING PROCEDURE FOR FROZEN CELLS

- Initiate culture as soon as possible upon receipt.

- Thaw by rapid agitation in 37°C water bath. Thawing should be rapid (within
40-60 seconds). As soon as the ice is melted, remove the ampule from the water
bath and immerse in 70% ethanol at room temperature. All of the operations from
this point on should be carried out under strict aseptic conditions.

- The cells are supplied in two different types of glass ampules. One is a
standard ampule, the neck of which must be scored with a sharp file that has
been immersed in ethanol. A definitive sharp nick about 1/8" in length on one
side is necessary. The second type is prescored and is identifiable by a gold
band around the ampule neck, and should not be scored with a file.

- Break the neck of the ampule between several folds of a sterile towel.

- Transfer the cell suspension and dilute it with the recommended culture
medium in a culture flask (see specific batch information above for dilution
ratio); incubate at 37°C with 5% CO₂ in air atmosphere. Since it is important
to avoid excessive alkalinity of the medium during recovery of the cells, it is
suggested that the culture medium be placed into the culture flask, tube, etc.
and the pH be adjusted, as necessary, prior to the addition of the ampule
contents. Note that the bicarbonate content of the culture medium will
determine whether an atmosphere containing CO₂ will be required.

- It is not necessary to remove the freezing additive. However, if desired, the
culture medium may be changed to remove the protective freezing additive
(dimethylsulfoxide) 24 hours after thawing. If it is desired that the freezing
additive be removed immediately, or that a more concentrated cell suspension be
obtained, centrifuge the above diluted suspension at approximately 125 x g for
10 minutes, discard the fluid and resuspend the cells with growth medium at the
dilution ratio given in the specific batch information above.

SUBCULTURE PROCEDURE

CHESEBRO PROTOCOL FOR GROWTH OF MOUSE LEUKEMIA LINES

1) MEDIUM RPMI 1640 with 200 units/ml penicillin. Before use add 10%
heat-inactivated fetal bovine serum and also 2-mercaptoethanol to a final
concentration of 10(-5) molar. (A stock of 0.01 M 2-ME solution in PBS which is
diluted 1:1000 into the tissue culture medium. The 2-ME does not need to be
fresh; and the stock solution has been used for 1-2 years.) Most, but not all,
of the mouse leukemia lines grow only in medium with 2-ME. MEM or McCoy's have
not been substituted for 1640 but may work. Most lines do not grow as well in
HEPES-containing medium.

2) Cells are grown in stationary suspension cultures at 37°C in 5% CO₂-95%
air. Use 25 cm² flasks containing 7 ml medium. Cells received after
shipping in tissue culture flasks full of medium should be pelleted by low
speed centrifugation, resuspended in fresh medium, and viable cells counted.
Initially, new cultures should be started at several different cell
concentrations (i.e. 3 x 10(4), 1 x 10(5), 3 x 10(5), 1 x 10(6) cells/ml) to
insure successful growth. This is particularly important if there is a high
proportion of dead cells in the population. Cells can also be started by I.V.
or I.P. injection of appropriate mouse strains (see #4 and #6 below).

3) For routine passage 1-7 x 10(5) cells from a fully grown-out culture are
seeded into a fresh flask containing 7 ml medium. Most lines grow to a maximum
final concentration of 1-4 x 10(6)/ml, it is usually not necessary to count
cells, but examine the cultures to see that the cells are in good condition
and present in high concentrations. Then 0.05 ml and 0.03 ml of suspension
culture are passed to two new flasks. The higher seeding inoculum usually
grows out to maximum concentration in 2-3 days, and the lower one in 4-5 days.
Under an inverted microscope at 400X, the viable-growing cells appear to have
a sharp even change in refractive index at the plasma membrane, and many
doublet cells are present. When the cells achieve their maximum concentration,
they begin to die off, the dead cells having a coarse granular appearance, the
nuclear appears obvious and clumped, the plasma membrane often jagged. The
ratio of dead to live cells increases rapidly in the next 1-2 days. Cultures
should be passed before the number of dead cells begins to increase. However,
in emergency situations, cultures with as few as 1% viable cells have been
passaged and saved.

4) Cells derived from ascites passaged lines can be passed in vivo in
appropriate mouse strains. Inject 0.5-5 x 10(6) cells I.P., however in few
cases where this has been checked, as few as 10(3) cells grew out reliably.
Mice may or may not get grossly enlarged abdomens. Most lines, mice will die
in 10-14 days. When unfamiliar with a particular line, cells should be
harvested from the peritoneal cavity as soon as any abdominal enlargement is
noted; since, with many lines, the transition from slight enlargement to death
can occur in 24 hours. Typical yield is about 500 x 10(6) cells/mouse.

5) All cell lines from both in vivo and in vitro sources have been frozen
successfully using standard techniques in medium plus 10% DMSO at
concentrations ranging from 1-50 x 10(6) cells/ml. (The higher the better in
terms of recovering more viable cells after thawing.) The FV lines which have
never been adapted to in vivo growth outside the lymphoid system are the most
difficult to freeze successfully. For these, freeze at a minimum concentration
of 10-15 x 10(6) cells/ml. Under these conditions, some batches work and some
do not.

6) Applicable to BB88 cells only (see TIB 55). Lines derived from splenic or
lymph node tumors which have not been adapted to growth outside the lymphoid
system can also be grown in vitro. Inject 5-30 x 10(6) cells I.V. Mice are
followed for splenic enlargement by palpation under ether anesthesia. Mice
with enlarged spleens or large abdominal masses are sacrificed and tumor-
containing organs are dissociated in buffer for passage or use.

Subculturing procedure

Medium Renewal: Every 2 to 3 days

Cultures can be maintained by addition or replacement of fresh medium.

Start cultures at 2 X 10 exp5 cells/ml and maintain between 1 X 10 exp5 and 1 X 10 exp6 cells/ml.

The cells may also be grown as ascites in syngeneic mice.

Inject 2 X 10 exp5 cell per mouse and harvest as soon as abdominal enlargement is observed (about 10 days).

Quality control specifications

Mycoplasma contamination: Not detected

History

Deposited as: Mus musculus
Depositors: B Chesebro
Special collection: Tumor Immunology Bank

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

References

Curated Citations

Chesebro B, et al. Lack of erythroid characteristics in Ia-positive leukemia cell lines induced by Friend murine leukemia virus: brief communication. J. Natl. Cancer Inst. 60: 239-242, 1978. PubMed: 564410

Chesebro B, et al. Characterization of Ia8 antigen, thy-1.2 antigen, complement receptors, and virus production in a group of murine virus-induced leukemia cell lines. J. Immunol. 117: 1267-1274, 1976. PubMed: 185295

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

Email

Send us a message

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