兔骨髓间充质干细胞 z-bio 2021-02-21 278

　　Objective: To study the effects of adenovirus (Ad) vector infection and bone morphogenetic protein 4BMP-4 overexpression on the proliferation and cartilage formation of rabbit bone marrow mesenchymal stem cells (BMSC) in vitro.

　　Methods: Using whole bone marrow to adhere, separate and obtain bone marrow mesenchymal stem cells, subculture to improve cell purity, and detect surface molecular markers by cell morphology and flow cytometry to identify cells, and establish the multiplicity of infection (MOI) )gradient. Group and reverse phase differential microscope were used to determine the best MOI by flow cytometry; the effect of BMP4 overexpression on enzyme-linked immunosorbent assay (ELISA) and collagen type II (COLL II) cell secretion. 9 (Sox9) and BMP-4; cell counting kit 8 (CCK-8) methods were used to identify the effect of BMP-4 overexpression on the proliferation of BMSCs in vitro.

　　Result: The primary cells are round, oval and spindle-shaped. The second-generation cells showed a typical fibroblast-like long spindle shape with a helical distribution and low expression of CD34 (1.94%) on the surface of hematopoietic stem cells. CD45 (2.13%), CD44 (98.2%), CD29 (99.7%) and CD90 (98.8%) are highly expressed. When the MOI is 100, the virus infection efficiency is high and the cell damage is minimal, which is the best MOI. Compared with the control group, the overexpression of BMP-4 significantly promoted the growth of BMSC, and may be secreted into the cartilage-specific indicators COLLII, BMP-4 and Sox9 (P\u003c0.05).

　　Conclusion: Overexpression of BMP4 may significantly promote the cartilage proliferation and differentiation of BMSC.