Method for simultaneous isolated culture of canine bone marrow mesenchymal stem cells and multifunctional hematopoietic stem cells
A technology of hematopoietic stem cells and bone marrow mesenchyme, which is applied in the direction of bone/connective tissue cells, animal cells, vertebrate cells, etc., can solve the problems of cumbersome operation steps, large cell damage, and many human factors, so as to overcome the screening steps, The effect of uniform cell shape and abundant material sources
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[0013] Example 1, step 1, in vitro separation of BMMSCs and HSCs: routinely anesthetized experimental dogs, skin preparation, disinfection, and draping of the posterior superior iliac spine, and a bone marrow puncture needle containing 1 ml of heparin sodium was used to extract a total of about 10 ml of bone marrow at 2 to 3 points , mix well to prevent coagulation, quickly transfer to a 50ml sterilized centrifuge tube, dilute and mix with an equal amount of DMEM / F12; take 10 centrifuge tubes with 5ml lymphocyte separation solution (1.077 g / ml) added to the bottom, and add carefully to the upper layer (paste Wall inflow) equal volume of bone marrow diluent, seal. Centrifuge at 2000 r / min at 4°C for 20 minutes to remove the upper fat cells, and carefully absorb the buffy coat cells at the middle junction. Mix and wash the interstitial cells with 10ml DMEM / F12 at 4°C, centrifuge at 2000r / min for 20min, remove the supernatant, and repeat the washing once; 5ml of DMEM / F12 medium ...
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