Amplification culture method for cloning adipose mesenchymal stem cells

Abstract

The invention discloses an amplification culture method for cloning adipose mesenchymal stem cells. The amplification culture method includes four steps of (1), extracting platelet-rich fibrin glue; (2), preparing special complete culture media; (3), carrying out isolated culture and subculture on primary adipose mesenchymal stem cells; (4), acquiring biological characteristics of different types of amplified adipose mesenchymal stem cells. The amplification culture method has the advantages that high-purity CD54 (+) adipose mesenchymal stem cell population can be obtained by the aid of the amplification culture method and is high in clone formation ability, self-renewal ability and adipogenic differentiation potential, novel methods can be provided for acquiring large quantities of stem seed cells in an in-vitro manner and applying the stem seed cells to regenerating adipose tissues, a novel treatment strategy can be provided for improving clinical repair for injury of soft tissue or congenital defects and abnormality of soft tissues of organs on body surfaces, and sufficient theoretical bases can be provided for guaranteeing the safety of clinical application of the adipose mesenchymal stem cells in the future.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, in particular to a method for expanding and culturing clones of adipose-derived mesenchymal stem cells. It relates to a high-efficiency expansion culture method of human adipose-derived mesenchymal stem cell clones and identification of related biological functions. Background technique [0002] Since Zuk et al first discovered in 2001 that adipose tissue contains mesenchymal stem cells with multidirectional differentiation potential, adipose-derived mesenchymal stem cells have gradually become one of the most promising seed cells in adult stem cells. Adipose tissue is easy to obtain through liposuction or surgery, and the trauma to the donor is relatively small, and it does not involve ethical issues. It has become an ideal tool in the application of cell therapy and tissue engineering. Many reports have confirmed that fat tissue Mesenchymal stem cells have broad clinical application prospe...

AI Technical Summary

Problems solved by technology

However, in the process of clinical application of adipose-derived mesenchymal stem cells, it is found that it is extremely difficult to obtain a large number of adipose-derived mesenchymal stem cells that can maintain the stable "stemness" of stem cells by using conventional culture techniques for clinical needs [Bunnell BA, Flaat M ,Gagliardi C,Patel B,Ripoll C.Adipose-derived stem cells:isolation,expansion and differentiation.Methods,2008,45(2):115-20.Cheng NC,Chen SY,Li JR,YoungTH.Short-term spheroid formation enhances the regenerative capacity of adipose-derived stem cells by promoting stemness, angiogenesis, and chemotaxis. Stem Cells Transl Med,2013,2(8):584-94.]

Images