KMS-21BM_ human multiple myeloma cells

KMS-21BM_Human multiple myeloma cells

Product Information

cell name：KMS-21BM_ human multiple myeloma cells

Species source: person

Gender and Age: woman67year

Organizational source: pleural effusion

Growth characteristics: Suspended growth

Cell morphology: Lymphocyte like

Cell specifications: 1 X 10⁶Cells/T25 or 1 mL cryovial

Cultivation conditions:RPMI 1640 + 10% fetal bovine serum，37 ℃, 5% CO2

Freezing conditions： 90% FBS + 10% DMSO

Passage method: 1：3 generations，2-3 days to pass down 1 generation

Cell culture operation

dry ice transportation：After receiving the cells, they should be immediately transferred to liquid nitrogen for storage, or-80°CShort term freezing or direct recovery of refrigerators

Room temperature transportation：After receiving the cells, please immediately remove the cell culture bottle from the packaging box and follow the steps below for culturing and passaging

subculturing：When the cell density reaches 80% -90%, passage culture can be carried out

Steps for operating cells in culture bottles

For adherent cultured cells, we will fill the entire culture bottle with culture medium before shipping to reduce the shedding of adherent cells during product transportation.

1. After receiving the cells, please pay attention to observe for contamination. Place the culture bottle under an inverted microscope and carefully inspect for turbidity and bacterial contamination. Due to the bumps during transportation and the sensitivity of some cells to temperature changes, there may be some cells shedding and floating. These cells are still alive and should not be discarded. They can be centrifuged and enriched before use.

2. For adherent cells, in a biosafety cabinet environment, use a vacuum pump to remove excess culture medium from the culture bottle until about 5-8 mL remains, and then select the appropriate CO2 content of 37 based on the culture medium℃Cultivate in a constant temperature incubator and loosen the bottle cap. If the cells have already filled the culture bottle, please subculture immediately.

3. For suspended cells, transfer the cells from the culture bottle to a centrifuge tube in a biosafety cabinet environment, centrifuge at 150 g for 5 minutes, remove the supernatant, blow off the cells with 5 mL of culture medium, transfer to a new culture bottle, and then place them in a suitable CO2 concentration of 37℃Cultivate in a constant temperature incubator.

Operation steps for freezing tube cells

Attention: To ensure high cell viability, please thaw and culture immediately after receiving the product.

1. Place the cryovial at 37℃Shake back and forth in the water bath to quickly thaw. To avoid pollution, ensure that the frozen storage pipe is placed above the water surface. Thawing should be done quickly, about 1 minute.

2. Once the liquid in the cryovial melts, immediately remove it and use alcohol to spray the surface of the cryovial. Starting from this step, subsequent operations must be completed in the biosafety cabinet.

3. Transfer the liquid from the cryovial to a centrifuge tube containing 5mL of complete culture medium, centrifuge at 150g for 5 minutes, and remove the supernatant using a vacuum pump.

4. Resuspend the cells after using the culture medium and transfer them to a new culture bottle. To ensure the survival rate of cell recovery, please incubate the culture medium at 37 ° C℃Preheat the water bath before use.

5. Place the cells in 37 cells containing appropriate CO2℃Cultivate in a constant temperature incubator.

Subculture of adherent cells

1. Suck and discard the culture medium from the culture bottle, and add PBSMoisturize once.

2. Join 1mL 0.25 (w/v) Trypsin-EDTASolution and placed in 37Incubate in a ℃ incubator until the cells detach from the wall. This process takes approximately 3-5minute

3. Join 2mLNeutralize trypsin in the culture medium and gently blow the cells off the surface of the culture bottle, dispersing the cells.

4. Collect the cell suspension15mLIn the centrifuge tube,Centrifuge at 150 g for 5 minutesUse a vacuum pump to remove the supernatant. Take an appropriate amount of culture medium to resuspend the cells and transfer them to a new culture bottle for cultivation.

Suspension cell passaging can refer to the following methods:

1、 Direct passage method

1. When the suspended cells reach 80% to 90% growth (yellowing of the cell suspension), they can be passaged;

2. usestrawDiscard cell suspension 1/2~1/3;

3. Add an appropriate amount of fresh culture medium and continue to cultivate.

   2、 Centrifugal passaging method (in the presence of cell debris)

4. Transfer the cell suspension tocentrifuge tubeInternally;

5. Centrifuge at 150 g for 5 minutes, discard the upper clear liquid;

6. Resuspend cells using fresh culture medium;

7. Suck an appropriate amount of cell suspension with a straw and fill it with a new oneculture flaskAdd an appropriate amount of fresh culture medium and continue to cultivate.

Cell cryopreservation operation

1. Centrifuge at 1000rpm for 5 minutes to remove the supernatant. Add 1 mL of serum to resuspend cells, add serum and DMSO according to the number of cells, and gently mix well，DMSO final concentration of 10%, cell density not less than 1 x 10⁶/mL， Each cryovial is used to freeze 1mL of cell suspension, and attention should be paid to labeling the cryovials properly；

2. Place the cryovial in a program cooling box and store it in a -80 ° C freezer for at least 2 hours before transferring it to liquid nitrogen storage.

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