It's So Simple To Eliminate Cell Clusters

When Single Suspension Cells Are Cultured In Growth Medium, It Is Not Uncommon To See Cell Loss In The Sample. When Cells Break Down, They Release DNA And Debris, Causing Cells To Clump Together, Making It Difficult For Them To Expand. Cell Aggregation Can Cause Both Apoptosis And Cell Death& Nbsp; As More Cells Die And Debris Is Released, The Problem Will Continue To Worsen. Handling Or Avoiding Cell Clustering As Soon As Possible Will Benefit The Subsequent Results Of The Experiment& Nbsp; Why Is Cell Clustering A Problem& Nbsp; The Objective Of Single Cell Suspension Medium Is To Provide Target Cells With As Many Growth Resources As Possible. This Includes Adequate Growth Nutrition And Sufficient Breeding Space. When Cells Gather Together, They Will Restrict Each Other, Reduce The Nutrient Throughput Of Cells, And Ultimately Affect The Growth Results& Nbsp; Cell Clustering Will Also Reduce The Recovery Rate Of Target Cells And Interfere With Labeling, Which Is Also The Reason For The Low Success Rate Of Suspension Cell Labeling. Some Cell Analysis Methods, Such As Flow Cytometry, Need To Properly Separate Cells. In Flow Cytometry, A Large Number Of Cells Pass Through The Machine In A Rapid Flow. This Machine, Called The Cytometer, Detects The Differences In Physical Properties Between Different Cells And Uses Fluorescent Lamps To Mark Them Accordingly. If Cells Gather Together When Passing Through The Test Tube, It Will Be Difficult For The Cell Analyzer To Accurately Measure Their Characteristics. This Will Cause Them To Be Incorrectly Classified And Have A Negative Impact On The Experimental Results. Reasons For Cell Clustering; One Of The Best Ways To Prevent Cell Aggregation Is To Understand What Causes Cell Aggregation And Take Preventive Measures To Avoid These Events. Some Reasons For Possible Cell Aggregation In Cultures Include:; 1. Overdigestion - Some Enzymes Used For Tissue Decomposition, Such As Trypsin, May Cause Cell Clotting If Overused; 2. Environmental Pressure - Physical Pressure And Repeated Temperature Changes Will Lead To Cell Death, Which Will Lead To The "sticky" DNA Molecules Of Cells Connecting Adjacent Cells Together; 3. Tissue Decomposition - When Preparing A Single Cell Suspension, The Use Of Enzyme, Mechanical Or Chemical Decomposition Strategies Can Cause Cells To Break Down, Leading To The Aggregation Of Debris& Nbsp; 4. Overgrowth - When Cells Are Too Dense In Their Culture Medium, Cells Will Start To Dissolve And Release Fragments, And The Sticky Factors In The Fragments Will Gather Cells Together; 5. Contamination - Some Bacterial And Fungal Pathogens Cause Cell Lysis; Agglomeration Is Usually A Sign Of Contamination Of Cell Culture  6. Although Some Of Them Occur Due To Experimental Errors, In Fact, Some Cells Have A Natural Clustering Phenomenon, Such As Most Semi Suspended Cells, BV-2 , NCI-H716 And So On. Generally, Semi Suspended Cells Need To Be Subcultured At About 70% Of The Overall Confluence, Which Can Reduce The Risk Of Cell Aggregation& Nbsp; How To Reduce Cell Clustering& Nbsp; 1.  Before Starting The Experiment, Some Measures Can Be Taken To Prevent Cell Aggregation. For Example, An Endonuclease Called DNase I Can Be Mixed Into The Culture Medium To Separate DNA From The Broken Cells. Decomposing These Superfluous Fragments Will Help Maintain The Space For Target Cells To Grow. However, Excessive Use Of Dnase I Will Lead To Cell Mutation, Which Will Eventually Affect Your Experimental Results. Therefore, It Is Not Necessary To Use It& Nbsp; 2.  Agglomeration Can Also Be Reduced Through Proper Handling And Use Of Equipment. If A Centrifuge Is Used To Separate Or Mix Samples, Setting It To The Correct Speed Can Reduce The Clustering. Generally, Cells Can Be Dispersed At A Faster Speed, But The Vulnerability Of Cells Must Also Be Considered At A High Speed. Therefore, Appropriately Extending The Centrifugation Time Is Also A Reliable Method Of Centrifuging And Suspending The Clustering Cells  3. There Are Also Some Methods To Separate Cells, Which Will Not Cause Excessive Damage If They Are Gathered Together. Certain Chemicals Called Chelating Agents Have An Affinity For Positively Charged Ions. According To The Nature Of Cell Clusters, Chelating Agents Can Be Added To Dissolve The Adhesion Between Them Without Damaging Cells. Ethylenediaminetetraacetic Acid (EDTA) Is A Chelating Agent Commonly Used To Dissolve Calcium Adhesion& Nbsp; 4. Another Method Of Cell Elimination Is Tritation. The Tritation Process Involves A Mild, Repeated Pipette To Break The Weak Adhesion Between Cells.