Method for separating and cultivating porcine marrow endothelial progenitor cell

Abstract

The invention relates to the biological cell technology field, particularly a method for separating and culturing miniature swine bone marrow-derived endothelial progenitor cells. The conventional EPCs culture method has the disadvantages of complexity and greatly reduced cell harvest rate by using immunomagnetic bead separation. The method in the invention comprises the steps of directly using myelomonocyte to conduct culture in vitro to obtain EPCs, adding growth factors in culture solution, planting in a definite density in a culture flask / vessel or a glass sheet enveloped by fibronectin,and conducing appropriate culture to obtain EPC. The method with high repeatability simplifies the steps of the conventional culture method, effectively avoids unnecessary cell loss caused by the conventional separation and culture method, increases the pick-up rate of EPCs, and saves a large amount of funds. The method of the invention also prepares for clinical application in future, and can reduce the usage of bone marrow and provide a technology platform for curing traumatic or ischemic diseases by autotransfusion.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a method for isolating and culturing porcine bone marrow endothelial progenitor cells. Background technique [0002] With the rapid development of stem cell technology and theory since the end of the 20th century, the research and clinical treatment of trauma-induced ischemic diseases and trauma repair mechanisms are facing new opportunities. Studies have shown that after body injury, stem / progenitor cells with multidirectional differentiation potential in the blood circulation can mobilize, proliferate, and migrate into injured organs under the action of cytokines / growth factors produced by inflammatory cells, and differentiate into corresponding tissue cells for further development. Injury repair, on the other hand, stem cells migrating to the injury site can also respond physiologically to different local stimuli (including hypoxia or ischemia), and sequentially release vasoactive...

AI Technical Summary

Problems solved by technology

[0012] The traditional culture method is more cumbersome, in the use of CD34 + MNC CD34 is unavoidable in the process of sorting MNC by antibody-coated immunomagnetic beads - The loss of the magnetic beads and the damage to the cells by the magnetic beads themselves will greatly reduce the acquisition rate of the cells. At the same time, the immunomagnetic bead separation method is expensive, especially as George et al. proposed the CD34 - After the concept of EPC, it was proved that the phenotypic expression rates on EPC were inconsistent, and the method of using immunomagnetic beads to sort MNC to obtain EPC has not been adopted.

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