Method for separating and cultivating porcine marrow endothelial progenitor cell
A technology of endothelial progenitor cells and culture methods, which is applied in the field of isolation and culture of porcine bone marrow endothelial progenitor cells, can solve the problems of high cost of immunomagnetic bead separation methods, inconsistent phenotypic expression rates, and reduced cell acquisition rate, etc. The effect of high efficiency, convenient material collection and wide source
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[0075]1) Isolation of mini-pig bone marrow mononuclear cells: Take 5-10 ml of anti-coagulated bone marrow from the ilium of mini-pig under sterile conditions, transfer the bone marrow cells into a test tube and mix them with sterile PBS solution 1:1 and centrifuge (20°C, 400g, 10 minute). After centrifugation, remove the supernatant, and then mix with sterile PBS solution 1:1. Take 10ml of Ficoll solution (Histopaque-10771, 1.077g / ml, Sigma company) and add it into a blank test tube in two parts; slowly drop the mixed bone marrow cell liquid into the test tube containing Ficoll solution (this step must be slow, be careful not to Break the plane between Ficoll and bone marrow cell fluid) and then centrifuge (20°C, 900g, 30 minutes). After centrifugation, first remove the uppermost supernatant (be careful not to take away the mononuclear cell layer under the supernatant), take out the mononuclear cell layer and add it to a blank test tube, then add 1:1 sterile PBS solution and ...
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