The Small Details Of Cell Resuscitation Make Your Cells Happy

The Holiday Ended Too Soon, And It Was Time To Start The Experiment Of Reviving Cells. Here Are Some Notes On Reviving Cells: 1; Cells Are Stored In Liquid Nitrogen At A Temperature Of - 196 ℃. Theoretically, The Storage Time Is Unlimited. As Long As There Is No Problem In Liquid Nitrogen And Cryopreservation, There Will Be No Major Problem In The Recovery. There Is No Expiry Date. What May Happen Is That The Longer The Cells Are Preserved, The More Advanced The Generation. The Basic Principle Of Cell Cryopreservation And Recovery Is Slow Freezing And Fast Melting. Resuscitation Must Be As Fast As Possible. Experiments Have Proved That This Can Maximize The Preservation Of Cell Vitality. 2.  Protective Measures Shall Be Taken When Taking Cells, And Antifreeze Gloves And Goggles Shall Be Worn. The Cell Cryopreservation Tube May Leak Into Liquid Nitrogen, And The Temperature In The Cryopreservation Tube Will Rise Sharply During Thawing, Which May Lead To Explosion. 3.  The Frozen Solution Must Be Completely Melted Within 1-2min. If The Recovery Temperature Is Too Slow, Cells Will Be Damaged. If The Wall Of The Cryopreservation Tube Is Thick And The Heat Insulation Effect Is Good, The Water Bath Temperature Can Be Appropriately Increased (37 ℃~40 ℃). It Is Recommended To Use Imported Products For The Cryopreservation Solution. For Example, When The Cell Systems' Primary Cell Cryopreservation Solution (product Number: 4Z0-705) Recovers, The Cell Survival Rate Is High, Or Shanghai Qida Biological Cell Seed Preservation Solution (product Number: SD0001) Can Be Selected. The DMSO Content Is 5%, And Trehalose, Propylene Glycol And Cell Growth Factor Are Added. 4; Book The Super Clean Table In Advance To Reduce The Waiting Time In The Ice Box After Recovery. If It Takes A Long Time (> 1h) To Wait After Resuscitation, It Is Recommended To Add More Than 15ml Of Medium To The Centrifuge Tube And Shake It Up To Dilute The Toxic Effect Of DMSO On Cells. For The Method Of Directly Adding Culture Medium Without Centrifugation, The Amount Of Medium Added Is A Problem To Be Considered. It Mainly Involves The Concentration Of DMSO. Generally, Cells With DMSO Content Of 5% Will Not Be Affected In 7.5ml Complete Medium, And Cells Can Grow Normally (this Is Very Useful). 5.  Whether Centrifugation Is Required After Cell Dissolution Depends On Specific Cell Conditions. There Are Two Main Ways. One Is That The Thawed Cell Suspension Is Directly Blown Evenly And Then Sub Packed Into Culture Bottles For Culture, And The Solution Is Changed The Next Day (the Solution Is Changed After The Wall Is Attached). Because The Purpose Of Centrifugation Is To Remove DMSO And Dead Cells, Which Is A Standard Process. But For Ordinary People, They Can't Grasp The Speed And Time Of Centrifugation. If They Don't Turn Enough, The Living Cells Will Be Thrown Away. After Turning, The Living Cells Will Be Pressed Too Much And Die. In Addition, It Is Easy To Be Polluted During Operation. The Other Is To Pour The Frozen Solution Clean After Centrifugation. Suspension Cells And Cells That Adhere To The Wall Slowly Must Be Centrifuged, Such As NCI-H716, LNCAP And Other Cells With Special Culture Methods. 6.  Some Cells Do Not Recover Until A Week After Recovery. Do Not Be Patient, Do Not Change The Fluid, Wait Patiently, And Make A Decision After Two Weeks. After Three Or Four Days Of Resuscitation, Do Not Discard All The Cells Without Any Movement. This Is The Problem With MLO-Y4. After The Technical Recovery, All Kinds Of Fancy Preferential Treatment For MLO-Y4 Did Not Last Long. Then Everyone Ignored It And Took It Out For More Than A Week, But It Grew Full.