Research Routine Of Tumor Stem Cells - Operation Steps Of Stem Cell Spheroidization Experiment

Tumor Stem Cells Are A Group Of Cells With Both Self-renewal Ability And Strong Invasion And Migration, Which Play An Important Role In Tumor Survival, Proliferation, Metastasis And Recurrence. The Tumor Cell Balling Experiment (size And Number Of Balling) Is The Gold Standard To Measure The Tumor Cell Dryness. In Tumor Stem Cell Spheroidization Experiment, The Selected Stem Cells Are Generally Cultured In Serum-free Medium And Ultra-low Adhesion Culture Dish, But They Can Also Be Cultured In Cell Lines. Cell Lines With A High Degree Of Malignancy Are Easy To Become Spheroids, But Some Cell Lines Are Very Difficult, So It Is Generally Recommended To Use Tumor Stem Cells For Culture. Cells Are Easy To Become Spheroids& Nbsp; (1) What Size Of Petri Dish, 6-hole Plate? 1. Petri Dishes Can Be 6, 12, 24 Hole Plates Or 100 Mm Petri Dishes; (2) Inoculation Density? 10000~100000 Pieces/ml, Or Do We Have To Find Out By Ourselves? Is There A Reference Density? 2. Density Needs To Be Explored By Yourself. Low Density Is Often Used For The Extraction Of Primary Tumor Stem Cells To Screen Cells That Can Grow Without Adhesion. High Density Is Used For The Expansion Of Cell Spheres. Generally Speaking, More Than 5000 Cells/ml Are Good For Spherulation. (3) How To Change The Liquid? 3. Add Complete Medium Twice A Week. Take 6-well Plate As An Example. Add 500ul Each Time And Shake Gently After Adding. Cell Spheres Grow Slowly. Growth Factors In The Medium Can Support At Least 2 Weeks. Direct Addition Of Tumor Stem Cell Serum-free Complete Culture Medium, Which Does Not Recommend Direct Addition Of Cell Growth Factors, Will Indeed Affect The Quality Of The Pellets. Once The Pellets Are Formed, They Need To Be Digested And Passaged. The Cell Ball Passaged Requires The Use Of Weak Digestive Enzymes, And TrypLE Express Can Be Used. It Is Said That Accutase And Dispase Can Also Be Used. Digestion Steps Are As Follows: 00001.  Seventy μ M Cell Sieve Was Used To Filter And Collect Cell Spheres For Trypsin Digestion. 00002.  2% BSA Solution Terminated Digestion And PBS Washed Cells Twice. 00003.  Resuspended Cells Were Counted. 00004.  Cells Were Cultured With Ultra-low Adsorption Cell Culture Plate (about 1000 Cells Were Added To Each Well In The 6-well Plate). 00005.  After About 10-14 Days Of Culture, Observe The Cell Spheroidization And Calculate SPF. Balling Ability Is An Important Method To Identify Tumor Stem Cells In Vitro. It Judges The Ability Of A Single Cell To Self Renew In An Appropriate Conditioned Medium, Which Is Generally Expressed By The Sphere Formation Efficiency (SFE). Calculation Method Of SFE: SFE=number Of Cell Spheres With Diameter Greater Than 75um In Each Hole/total Number Of Original Inoculated Cells In Each Hole Click To Go Directly To The Official Website Of Shanghai Qida Biology. The Tumor Stem Cell Serum Free Culture Medium Kit Is Added With SMOC-2 To Promote Tumor Stem Cell Spheroidization, And Contains The Protein Required For Stem Cell Growth, To Maintain Stem Cell Growth And Spheroidization, And To Escort Your Spheroidization Experiment.