Operating Procedures For Mycoplasma Clearance By Animal Adoption

The Cells Grow And Become Slower And Slower. Then You Can Add Whatever You Want. They Will Be There. If They Don't Grow, They Will Even Roll Their Eyes To Show You That They May Be Infected With Mycoplasma. Try Our New Recommended Method Of Mycoplasma Adoption For Small Animals. The Adoptive Methods Are As Follows: 1. Three 4-5 Week Old BALB/c Nude Mice Were Taken, And Each Of Them Was Subcutaneously Inoculated With Tumor Forming Cells Under The Right Armpit. When The Average Tumor Volume Was About 1.0 X 1.0 X 1.0 Cm3, Tumor Bearing Mice Were Obtained. The Tumor Bearing Mice Were Killed For Cervical Dislocation And Immersed In 75% (volume Percentage Concentration) Ethanol Water Solution For 3-5 Min To Obtain The Sterilized Mice; In The Ultra Clean Workbench, Use Sterile Scissors And Sterile Tweezers To Cut The Mouse Skin From The Sterilized Mouse Chest, And Carefully Peel Off The Tumor Tissue. Slightly Cut The Tumor Tissue With Ophthalmic Scissors (at This Time, The Tumor Tissue Size Is 0.5 X 0.5 X 0.5cm3). 2. Soak It In 0.01mol/L PBS (to Make The Tumor Tissue Completely Infiltrate Into 0.01mol/L PBS) For 2min, And Then Take It Out To Obtain The Tumor Tissue After Soaking. Use An Ophthalmic Scissors To Cut The Soaked Tumor Tissue On A Sterile Plate (the Size Of The Tumor Tissue At This Time Is 0.2X0.2X0.2mm3). In The Process Of Cutting The Soaked Tumor Tissue, Add 0.0 Imo I/L PBS At The Right Time To Keep The Tumor Tissue Moist. Add Collagenase I Solution To The Sterile Plate Containing The Cut Tumor Tissue, Blow Away The Tumor Tissue With An I ML Gun Head (it Is Recommended To Cut Off The Gun Head For 3mm To Better Blow Away The Cells) To Obtain The Collagenase Tumor Tissue Mixture. 3. Transfer The Collagenase Tumor Tissue Mixture To A 50mL Centrifuge Tube, And Shake The 50mL Centrifuge Tube Containing The Collagenase Tumor Tissue Mixture On The Oscillator For 3min To Obtain The Shocked Collagenase Tumor Tissue Mixture. Incubate The Shocked Collagenase Tumor Tissue Mixture In A 37 ° C Water Bath For 2min To Obtain The Collagenase Digested Tumor Tissue, Then Centrifugation Washing Centrifugation Washing Operation Steps Are As Follows: Centrifugate The Tumor Tissue Digested By Incubated Collagenase At 300g For 6 Min, Discard The Supernatant, Add 20 ML 0.01mol/L PBS, Centrifugate At 300g For 6 Min After Blowing Evenly, Discard The Supernatant, Add 20 ML 0.01mol/L PBS, Centrifugate At 300g For 6 Min After Blowing Evenly, Discard The Supernatant, And Obtain The Tumor Tissue Digested By Collagenase Three Times. Add The Complete Culture Medium Of The Cell, And Culture It In The Cell Incubator For Two Days. The Volume Percentage Of CO2 In The Cell Incubator Is 6-8%, And The Temperature Is 35-38 ° C. The Tumor Tissue With Loose Tissue Structure Is Obtained, And Then Add Collagenase Solution To The Tumor Tissue With Loose Structure For Digestion. Repeat The Previous Centrifugation Procedure, Add Pancreatin Solution To The Tumor Tissue With Loose Tissue Structure To Obtain The Pancreatin Tumor Tissue Mixture. After Placing The Pancreatin Tumor Tissue Mixture On The Ultra Clean Workbench At Room Temperature For 1-3 Minutes, Add Double Fetal Bovine Serum FBS (Catalog: SD0200) To Terminate Digestion, And Pour Out The Pancreatin Tumor Tissue Suspension To Obtain The Tumor Single Cells That Adhere To The Wall After Pancreatin Digestion. Add Complete Cell Culture Medium And Place It In The Incubator For Culture. After Passage, It Is Sent To STR For Detection And Mycoplasma Detection. This Operation Can Remove Mycoplasma Contamination From Most Cells And Improve Cell Vitality. Friendly Tips, The Operation Method Of Less Than 1000 Words Seems Simple, But It May Take 1-2 Months To Operate. In Addition, The Separation Of Primary Tumor Cells Is Also Very Easy To Use. If You Have Resources, You Can Try It And Communicate With Each Other.