Prevention And Treatment Of Mycoplasma "Trouble Sperm" In Cell Culture

Mycoplasma Is A Common Contamination In Cell Culture,. As Mycoplasma Belongs To The Smallest Prokaryote With A Diameter Of 0.2-2 µ M, And Its Morphology Is Changeable, It Will "haunt" The Infected Medium Even If It Is Filtered For 20 Times With A Filter Screen, And Because It Lacks Cell Wall, It Can Successfully Escape The "pursuit" Of Most Antibiotics Without This Coat. In The Cell Culture Medium, Mycoplasma Can Reach 107-108CFU/mL (CFU=Colony Forming Unit, Which Is A Measure Of The Concentration Of Mycoplasma) Without Making The Culture Medium Turbid And Affecting The Cell Growth. Because It Will Not Affect The Cells At The Initial Infection, It Is Estimated That The Kerria Melanogaster Will Be Impressed By Its Concealment. So What Effect Does Mycoplasma Pollution Have On Cells? 1. Mycoplasma Contamination Leads To Cell Contraction, Adherent Cells Shedding And Splitting, And Cell Vacuolization... 2. Causes Cell Protein, DNA, RRNA And MRNA; Synthesis Disorder Of; 3. Change The Composition Of The Culture Medium (for Example, Mycoplasma Fermentum Degrades Sugars To Produce Acidic Substances) And Affect Cell Metabolism 4. Destroy The Integrity Of Cell Membrane And Affect Cell Signal Transmission; 5. Cause Chromosome Abnormality Of Cells; 6. It Can Reduce The Carcinogenic Ability Of Malignant Cells, Affect The Differentiation Of Lymphocytes And The Production Of Viruses In Cells; What Are The Detection Methods For Mycoplasma Contamination? 1 Phase Contrast Microscope Detection; Observation Of Lichen Red Staining In Hypotonic Treatment; 3 DNA Fluorescence Staining; 4 Electron Microscopy; 5 3-H Thymine Incorporation Method; Polymerase Chain Reaction; 7 Immunological Methods; Culture Of Mycoplasma& Nbsp; Remedial Method For Mycoplasma Contamination: Antibiotic Elimination Method: You Can Use 5-10 Times Of The Commonly Used Amount Of Impact Method, Add High Concentrations Of Antibiotics, And Then Act For 24-48 Hours, And Then Change Into The Conventional Culture Medium. Sometimes It May Be Effective. Recommended Dosage: Gentamicin 200ug/ml, Tetracycline 10ug/ml, Kanamycin 50ug/ml. For The Use Of Antibiotics Contaminated With Mycoplasma, Preventive Use Is Better Than The Use After Contamination. 2 . Heat Sterilization: According To The Characteristics Of Poor Heat Resistance Of Mycoplasma. Mycoplasma Can Be Killed By Exposing Cells Contaminated With Mycoplasma To 41 Degrees For 5-10 Hours. However, As 41 ° C Also Has A Greater Impact On The Cultured Cells Themselves, A Pre-test Should Be Carried Out Before Treatment To Determine The Treatment Time For Maximum Killing Of Mycoplasma And Less Impact On Cells. 3. Adoptive Method In Nude Mice: The Cells Contaminated With Mycoplasma Were Digested With 0,25% Trypsin And 0.02% EDTA To Form A Single Cell Suspension, Washed And Centrifuged Twice With PBS, And The Cell Density Was Adjusted To 10 ^ 7/ml; After Subcutaneous Inoculation Of 0.2ml Cells On The Back Of Mice For 10-15 Days, The Transplanted Tumor Was Formed; Three To Four Weeks Later, The Transplanted Tumor Tissue Was Taken Out Under Aseptic Conditions, And The Necrotic Part Was Removed. The Tissue Was Crushed Under A 200 Mesh Sieve With A Syringe Core To Release The Cells Into A Serum Free DMEM Culture Solution Containing 0.1% EDTA: The Cell Suspension Was Carefully Added To The Upper Layer Of The Centrifuge Tube Containing Lymphocyte Separator, And The Horizontal Centrifugation Was 3000 R/min, 20 Min At Room Temperature; Suck Out The Cells In The Interface Layer Of Two Kinds Of Liquids With A Straw, Place Them In The Serum Free DMEM Culture Solution, Wash And Centrifuge The Cells Once With The Serum Free DMEM, And Adjust The Cell Density To 5 With The Complete Culture Solution × 105 Pieces/ml, Placed In 37 ℃, 5% CO2 Incubator& Nbsp; Want To Know More About Cells? You Can Visit Our Official Website: Shanghai Qida Biotechnology Co., Ltd. Or Follow Us& Nbsp;