Stem cell preparation for preventing acute graft versus host disease
A technology of graft-versus-host and stem cell preparations, applied in the field of stem cells and immunology, can solve problems affecting the treatment effect of allogeneic hematopoietic stem cell transplantation, difficult comprehensive treatment schemes, unfavorable immune reconstitution, etc., to promote hematopoietic reconstitution and immune reconstitution, The effect of reducing the severity and reducing the incidence rate
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[0023] (1) Isolation, identification and passage expansion of mesenchymal stem cells
[0024] Before transplantation, allogeneic healthy bone marrow was collected, and mononuclear cells were harvested for culture. With the help of MSC's adherent characteristics, the medium was changed repeatedly and the suspended cells were discarded, so as to gradually obtain pure bone marrow mesenchymal stem cells. Through morphological observation, flow cytometry analysis, and multi-directional differentiation potential identification of osteogenic, adipogenic, and neural-like cell differentiation, it was confirmed that the isolated cells were mesenchymal stem cells, which were continuously passaged and expanded for future use.
[0025] (2) Preparation of splenocyte suspension
[0026] Take the spleen of the donor rat, cut it into small pieces with scissors, grind it gently with a syringe needle core on a 200-mesh steel mesh, and let it pass through to form a spleen cell suspension. Remove...
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