Extraction Of MSC Cells From Mice

MMSC Cells Generally Adopt The Procedures Of Long Bone Anatomy And Bone Marrow Isolation. The Overall Goal Is To Quickly And Systematically Remove Long Bones And Collect Bone Marrow Cells For Further Experimental Analysis. This Method Is Helpful To Answer The Key Questions About Bone Marrow Biology, Tumor Metastasis And Immunology. The Main Advantage Of This Technology Is That The Dissection Process Is Fast And Standardized, And The Bone Marrow Isolation Procedure Is Quite Sterile. First, The Mice Are Killed And Placed In The Supine Position. Then Fix The Four Paw Pads Of The Mice Under The Ankle Joint, Spray The Mice With 70% Ethanol, And Wash The Legs Thoroughly. Next, Make A Small Incision On The Right Side Of The Midline Of The Lower Abdomen Just Above The Hip, And Extend The Incision Down To The Leg, Through The Ankle Joint. Pull The Skin Back. Then Cut The Quadriceps Femoris Fixed At The Proximal End Of The Femur To Expose The Front Of The Leg Bone, And Fix The Muscle From The Leg With A Pin At A 45 Degree Angle With The Board. Using Scissors Against The Back Of The Femur, Cut The Hamstrings From The Knee Joint. Then Pull Back The Skin And Hamstring Muscle Fixed At The Proximal End Of The Femur To Expose The Back Side Of The Bone, And Fix The Hamstring Muscle From The Leg To Make The Fixing Pin Form A 45 Degree Angle& Nbsp; Grasp The Distal Femur With Tweezers, Just Above The Knee Joint. Then Guide The Scissors Blades On Both Sides Of The Femoral Shaft Towards The Hip Joint. After Reaching The Femoral Head, Twist The Scissors And Move The Top Blade Directly Above The Femoral Head To Dislocate The Femur& Nbsp; Grasp The Top Of The Femoral Shaft With Tweezers, Cut The Soft Tissue From The Femoral Head, And Take The Femur Out Of The Acetabulum. Then Pull The Whole Leg Bone, Including The Femur, Knee And Tibia, Up And Away From The Body, And Carefully Cut Off The Connective Tissue And Muscle Connecting The Leg With The Skin. When All The Connective Tissue Has Been Removed, Stretch The Ankle Joint And Twist The Scissors To Dislocate The Tibia& Nbsp; Grasp The Distal End Of The Tibia, Be Careful Not To Cut The Tendon, And Pull The Tibia Upward Away From The Body And Nail Plate. Then Remove Any Remaining Connective Tissue On The Long Bone Of The Knee, As Well As Any Muscle Or Connective Tissue Attached To The Femur And Tibia. Grasp The Femur So That The Patella Is Outward And The Femoral Head Is Downward& Nbsp; Excessive Extension Of The Knee Joint And Twisting Of The Scissors Cause Dislocation Of The Tibia And Femur. Then Remove All The Connective Tissue Connecting The Femur And Tibia. Next, Grasp The Femur, Make The Front Side Outward And The Femoral Head Downward, And Then Guide The Scissors Upward Along The Femoral Shaft To The Condyle& Nbsp; Gently Rotate The Scissors Back And Forth To Remove The Condyle, Patella And Epiphysis And Expose The Metaphysis. Then, Use Tweezers, Scissors And Low Dust Wiping Paper To Remove Any Muscle Or Connective Tissue Attached To The Femur. Next, Grasp The Tibia, With The Front Side Facing Outward And The Ankle End Facing Downward& Nbsp; If The Tibial Epiphysis Is Intact, Lead The Scissors Up To The Condyle Along The Tibial Axis, Turn The Scissors Back And Forth Slightly, Remove The Condyle And Epiphysis, And Expose The Tibial Epiphysis. Then Remove Any Muscle Or Connective Tissue Attached To The Tibia, As Shown. In Order To Obtain Bone Marrow, First Pass A No. 18 Needle Through The Bottom Of A 0.5ml Microcentrifuge Tube& Nbsp; Then Put The Long Bone Into The Tube, Knee Down, And Cover It. Place A 0.5 ML Microcentrifuge Tube Into A 1.5 ML Microcentrifuge Tube And Centrifuge For 15 Seconds In A Microcentrifuge Tube Greater Than Or Equal To 10000 Times G. Visually Check That The Bone Marrow Has Been Rotated Out Of The Bone& Nbsp;• Use A 10mL Syringe To Extract Bone Marrow Cells Transfer To A 50mL Tapered Pipe And Increase The Volume To 50mL With FSB Transfer 50uL Sample To Eppendorf Tube And Count Cells Rotate Cells At 1200 Rpm For 8 Minutes Pour The Supernatant From The Centrifuged Cells And Transfer It To The Complete Stem Cell Culture Medium For Culture & Nbsp;& Nbsp; & Nbsp; & Nbsp; & Nbsp; & Nbsp; & Nbsp;& Nbsp;& Nbsp;& Nbsp;& Nbsp;& Nbsp;& Nbsp;& Nbsp; The First Day Of Normal Culture& Nbsp;& Nbsp;& Nbsp;& Nbsp;& Nbsp;& Nbsp;& Nbsp;& Nbsp;& Nbsp;& Nbsp;& Nbsp;& Nbsp;& Nbsp;& Nbsp;& Nbsp;& Nbsp;& Nbsp;& Nbsp; MMSC Bm Of Mice Extracted On The Third Day Of Normal Culture& Nbsp;& Nbsp;& Nbsp;& Nbsp;& Nbsp;& Nbsp;& Nbsp;& Nbsp;& Nbsp;& Nbsp;& Nbsp; The Mesenchymal Stem Cell Complete Culture Medium (Catalog: P2001) Of Shanghai Qida Biotechnology Co., Ltd. Was Used For Purification And Culture.