Preparation method of tissue engineering cell sheet with osteogenic differentiation and vasculogenesis
An angiogenesis and tissue engineering technology, applied in the field of transplantation, can solve the problems of extracellular matrix damage, decreased cell activity, low cell shelf rate and low cell density, and achieves the effect of maintaining coordination, avoiding adverse reactions and improving signal transmission.
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[0010] The present invention is further described below in conjunction with specific embodiment, but protection scope of the present invention is not limited thereto:
[0011] The preparation method of the tissue engineering cell sheet with osteogenic differentiation and angiogenesis of the present invention comprises the following steps:
[0012] 1. Isolation of bone marrow mesenchymal stem cells (MSCs)
[0013] Healthy adult Sprague-Dawley rats were sacrificed after anesthesia, soaked in 75% alcohol for 10 min, and the long bones of the limbs were removed under aseptic conditions, the soft tissue attached to the bone surface was removed, and washed with PBS buffer. The metaphysis at both ends was resected with a rongeur to expose the bone marrow cavity. Thoroughly flush the bone marrow cavity with low-sugar DMEM medium with a volume concentration of 10% FBS, and then repeatedly flush the flushed bone marrow with a syringe with a No. 7 needle and a No. 5 needle to fully ...
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