Method for promoting directional osteogenic differentiation of mesenchymal stem cells
A technology of mesenchymal stem cells and osteogenic differentiation, applied in biochemical equipment and methods, animal cells, vertebrate cells, etc., can solve the problems of not being able to provide, and achieve good magnetoelectric coupling performance, best safety and controllability Effect
Pending Publication Date: 2021-01-01
PEKING UNIV SCHOOL OF STOMATOLOGY
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Problems solved by technology
[0005] Aiming at the technical problem that the existing implant materials cannot dynamically regulate the surface potential after implantation to regulate cell function and differentiation, the present invention provides an electromagnetic material that can be regulated after implantation, thereby inducing the polarization of macrophage M2 and promoting Methods of Directed Osteogenic Differentiation of Mesenchymal Cells
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Embodiment 1
[0030] (1) Expansion and culture of mouse macrophage cell line (macrophage Raw264.7): adopt high-glucose DMEM medium without induction factors, Raw264.7 cells are resuscitated and adhered to the T25 culture flask for culture, every 1- After 2 days of washing with PBS and replacing with new medium (high glucose DMEM medium), the adherent cells were subcultured by blowing gently.
[0031](2) Expansion culture of mesenchymal stem cells: using α-MEM medium without induction factors, mesenchymal stem cells (MSCs) were cultured adherently in T25 culture flasks after recovery, and 0.25% trypsin was used every 2-3 days Digestion for passaging.
[0032] (3) Induce M2 polarization of macrophages and directional osteogenic differentiation of mesenchymal stem cells: Inoculate Raw264.7 macrophages of passage 8-15 on charged biomimetic membrane materials, and inoculate MSCs of passages 3-5 in co-culture In a (transwell) chamber (Millipore, USA), an indirect co-culture system was constructe...
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The invention relates to a method for promoting directional osteogenic differentiation of mesenchymal stem cells, and solves the technical problems that an existing implant material cannot dynamicallyregulate and control surface potential in vitro after implantation to regulate cell functions and differentiation. The method comprises the following steps: carrying out amplification culture on macrophages; carrying out amplification culture on the mesenchymal stem cells; regulating and controlling macrophage polarization and osteogenic differentiation of the mesenchymal stem cells, wherein macrophages are inoculated to the surface of a charged bionic implant membrane material with a magnetoelectric coupling effect, and the mesenchymal stem cells are inoculated to a co-culture chamber; transferring the co-culture chamber inoculated with the mesenchymal stem cells into a charged bionic membrane material culture plate inoculated with the macrophages to establish an indirect co-culture system; and changing an external magnetic field to cause changes of a magnetoelectric microenvironment, and regulating and controlling macrophage polarization so as to promote osteogenic differentiation of the mesenchymal stem cells. The method can be used for promoting directional osteogenic differentiation of the mesenchymal stem cells.
Description
technical field [0001] The invention relates to a method for promoting cell differentiation, in particular to a method for promoting directional osteogenic differentiation of mesenchymal stem cells. Background technique [0002] With the economic development in recent years, the cases of bone defects caused by factors such as trauma and tumors have increased. At present, implantation is often used to repair bone defects. Implants can be divided into autologous bone, allogeneic bone and synthetic materials. Among them, autologous bone transplantation often has problems such as insufficient bone mass in the donor site and secondary infection in the operation site. There are fewer donors for allograft bone, and there are risks such as immune rejection. In order to solve the above problems, the use of artificial synthetic materials instead of bone tissue for implant repair is currently a research hotspot in tissue engineering. Researchers have discovered that a natural elect...
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Login to View More IPC IPC(8): C12N5/077C12N5/0775C12N5/0786C12N13/00
CPCC12N5/0654C12N5/0662C12N5/0645C12N13/00C12N2502/1157C12N2502/1352C12N2529/00
Inventor 邓旭亮张冯依刘雯雯
Owner PEKING UNIV SCHOOL OF STOMATOLOGY