A method for promoting osteogenic differentiation and inhibiting adipogenic differentiation of adipose stem cells by using sema3a gene

Abstract

The invention disclose a method using Sema3A genes to promote the osteogenic differentiation of adipose-derived stem cells and inhibit the adipogenic differentiation of the adipose-derived stem cells and belongs to the technical field of cell biology.The method has the advantages that recombinant plasmids containing Sema3A gene sequences are built, the Sema3A recombinant plasmids and auxiliary packaging plasmids are transfected into 293T cells to obtain Sema3A lentiviral vectors, the lentiviral vector introduces the Sema3A genes into the adipose-derived stem cells by the aid of polybrene, and accordingly the osteogenic differentiation of the adipose-derived stem cells is promoted evidently, and the adipogenic differentiation of the adipose-derived stem cells is inhibited at the same time; the adipose-derived stem cells can be well used for promoting bone defect healing and implants-osseointegration due to the fact that the Sema3A genes are introduced, and the advantages of wide source and easiness in obtaining of the adipose-derived stem cells are brought into full play.

Description

technical field [0001] The invention belongs to the technical field of cell biology, and specifically relates to a method for promoting osteogenic differentiation and inhibiting adipogenic differentiation of adipose stem cells by adopting Sema3A gene. Background technique [0002] Stem cells are a type of cells that have the ability to self-replicate and differentiate into various functional cells. It has good proliferation and immune regulation capabilities and the potential to regenerate various tissues and organs, and can significantly expand the number of cells involved in tissue repair. Researchers have successively developed stem cell technologies based on the multi-directional differentiation potential of stem cells, including bone marrow transplantation for hematopoietic Dysfunction, the stem cells in the human skin are separated and cultured to make artificial skin for burn wound treatment and so on. [0003] With the progress of development, large bone defects los...

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Problems solved by technology

[0004] At present, there are mainly four methods to regulate the differentiation of adipose stem cells: one is to use fluorescent labeling flow cytometry to select adipose stem cells with strong osteogenic ability from the stem cell population for expansion and then use them. One-time use of adipose stem cells requires fluorescent labeling and flow cytometry screening, which is time-consuming and laborious, which reduces the advantages of adipose-derived stem cells and ease of use; the second is to use osteogenic induction fluid to induce osteogenesis of adipose-derived stem cells, and then apply them to tissues To promote the healing of bone defects on engineered bone, this method only exerts the osteogenic ability of adipose-derived stem cells in vitro in advance, and does not essentially promote the osteogenic differentiation of adipose-derived stem cells, and has no effect on the adipogenic differentiation of adipose-derived stem cells; The third is to use BMP and other cytokines that can promote the osteogenic differentiation of adipose-derived stem cells to directly add to the culture medium or play a role on adipose-derived stem cells in vivo in various slow-release ways, which can essentially improve the osteogenesis of adipose-derived stem cells differentiation ability, but its action time is limited, and it is difficult to maintain a stable and long-lasting effect of promoting osteogenesis or inhibiting adipogenesis; the fourth is to use small molecular compounds, such as dexamethasone and statins, to intervene in adipose-derived stem cells to improve their osteogenic ability. This method is relatively simple and less expensive, but the effect is limited

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