LK 35.2小鼠淋巴瘤细胞B细胞HB-98 BioVector NTCC质粒载体菌种细胞基因保藏中心

LK 35.2小鼠淋巴瘤细胞B细胞HB-98

BioVector NTCC质粒载体菌种细胞基因保藏中心  

Organism: Mus musculus (B cell); Mus musculus (lymphoma), mouse (B cell); mouse (lymphoma)

Disease: Lymphoma

Cell Type: hybridoma: B lymphocyte

Morphology: lymphoblast

Growth Properties: suspension

Complete Growth Medium

The base medium for this cell line is Dulbecco's Modified Eagle's Medium.

To make the complete growth medium, add the

following components to the base medium: fetal bovine

serum to a final concentration of 10%.

HANDLING PROCEDURE FOR FROZEN CELLS

1. Initiate culture as soon as possible upon receipt.

2. Thaw by rapid agitation in 37°C water bath. Thawing should be rapid (within 4060

seconds).

As soon as the ice is melted, remove the ampule from the water bath. All of the operations

from this point on should be carried out under strict aseptic conditions.

3. 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 vial

contents. Note that the bicarbonate content of the culture medium will determine whether an

atmosphere containing CO2 will be required.

4. 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 xg for 10 minutes, discard the fluid and resuspend the

cells with growth medium at the dilution ratio given in the specific batch information above.

HANDLING PROCEDURE FOR FLASK CULTURES (SUSPENSION)

The flask was seeded with cells (see specific batch information above for

concentration), grown and completely filled with medium to prevent loss of

cells in transit. Upon receipt incubate the flask in an upright position for

several hours to return the flask contents to 37°C. After the temperature has

equilibrated, aseptically remove the entire contents of the flask and

centrifuge at 300 xg for 15 minutes. Resuspend the cell pellet in 1012

mL of

the shipping medium. From this suspension remove a sample for a cell count and

viability so that the cell density of the suspension can be adjusted to

24

X 105 viable cells/mL. If the suspension needs to be diluted use the shipping medium.

Incubate the culture in a flat position at 37°C in a 5% CO2 in air atmosphere. Maintain the cell

density of the culture as suggested under the

subculture procedure described above.

Medium Renewal: Every 2 to 3 days

Cultures can be maintained by addition or replacement of fresh medium. Start cultures at 4 X 10

exp5 cells/ml and maintain between 3 X 10 exp5 and 2 X 10 exp6 cells/ml.

Comments：

The LK 35.2 bears surface IAd,k and IEd, k molecules and can present antigen to appropriate I

region restricted T cell hybridomas.

The line was produced by fusing A20.2J lymphoma cells (IAd,

IEd) with T cell depleted spleen cells from B10.BR mice (IAk,IEk).

Tested and found negative for ectromelia virus (mousepox).

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