C57BL/6 cell specific culture medium

C57BL/6 cell specific culture medium

C57BL/6The cell culture medium has been carefully optimized by the team, and after long-term testing, this product can maintainC57BL/6Excellent growth state of cells.

This product already includesC57BL/6The various components required for cell growth can be directly used without adding any ingredientsC57BL/6Cell culture.

Main ingredients of the product:

DMEMbasic medium 405mL

Defined FBS 75 mL

GlutaMAX-1 5 mL

MEM NEAANon essential amino acids 5 mL

Sodium Pyruvatesodium pyruvate 5 mL

P/S 5 mL

B-Mercaptoethanol 0.5 mL（1000X)

LIF 5Mg（10ng/mL）

Transportation and storage

Transportation: Low temperature transportation in an insulated box containing biological ice packs

保存方法： 2℃～8℃， 保存2Months;

1）RecoveryC57BL/6cell: will contain1mLThe freezing tube for cell suspension 37Quickly shake and thaw in a ℃ water bath, then add4mLMix the culture medium evenly. in1000RPMCentrifuge under conditions 4 Minute, discard the supernatant and add more1-2mLBlow the culture medium evenly. Then add all cell suspensions to the culture bottle and culture overnight (or add the cell suspensions)10cmIn the dish, add about8mlCulture medium, overnight cultivation). Change the liquid and check the next dayC57BL/6cellDensity.

2）Cell passage: IfC57BL/6cellDensity reaches80%-90%You can proceed with subculture.

1. Discard the culture supernatant and use one that does not contain calcium or magnesium ionsPBSrinseC57BL/6cell1-2Next time.

2. plus1mlDigestive fluid（0.25%Trypsin-0.53mM EDTA）In the culture bottle, place it 37Digestion in a ℃ incubator1-2Minutes, then observe the digestion of cells under a microscope, ifC57BL/6cellMost of them become round and fall off, quickly take them back to the workstation, lightly tap the culture bottle a few times, and add a small amount of culture medium to stop digestion.

3. press6-8ml/Add culture medium to the bottle, gently mix well, and then aspirate1000RPMCentrifuge under conditions 4 Minute, discard the supernatant and add more1-2mLBlow the culture medium evenly.

4. Press the cell suspension 1：2 Proportionally divided into new categories8mlIn a new dish or bottle of culture medium.

3）Cell cryopreservation: pendingC57BL/6cellWhen the growth state is good, cell cryopreservation can be performed. belowT25Bottles are classified;

1. After discarding the culture medium during cell cryopreservation,PBSAfter cleaning once, add1mlAfter the cells become round and shed, add1mlSerum containing culture medium can terminate digestion and can be counted using a hemocytometer.

2. 4 min 1000rpmCentrifuge to remove the supernatant. plus1mlSerum resuspended cells, add serum andDMSOGently mix well,DMSOThe final concentration is10%The cell density is not less than1x106/mlEach cryopreservation tube is frozen1mlCell suspension, pay attention to labeling the cryovials properly.

3. Place the cryovial in the program cooling box and put it in-80 The refrigerator,2 Transfer to liquid nitrogen storage after one hour. Record the location of the freezer for future retrieval.

1.useC57BL/6cellAttention should be paid to aseptic operation to avoid contamination.

2.This product contains serum and bispecific antibodies. If there is no special need, there is no need to add additional serum and bispecific antibodies, and they can be used directly.

3.To maintain thisC57BL/6cellIt has excellent usability and should not be placed in room temperature or high temperature environments for a long time.

4.All products should be used within the shelf life. If they exceed the shelf life, they must be discarded.

5.This product is only for further scientific research use and cannot be applied to clinical or other aspects.

6.After freezing and thawing of the culture medium, a small amount of flocculent material may precipitate, which does not affectC57BL/6cellNormal use.