In Vitro Biocompatibility Study On The Human Decellularized Adipose Tissue Combined With Adipose Derived Stem Cells

With the progress treatment of breast cancer,while obtaining the long-term survival rate,patients are particularly demanding on the physical beauty.Postoperative breast reconstruction of breast cancer have been widely accepted as a routine treatment.The development of tissue engineering provides a new way to solve this problem.It can be implanted in the body with a small amount of seed cells after being amplified in vitro with biomaterials,minimizing complications and effectively solving the problem of prosthetic contracture deformity,reducing the rate of secondary surgery.We speculate it will bring new hope for the reconstruction of breast tissue defects.Studies shows that ADSCs with the characteristics of easy separation,strong proliferation,multidirectional differentiation and low immunogenicity,effectively solve the problem of breast reconstruction safety.Stem cell-scaffold co-transplantation is an effective way to reduce cell damage and increase therapeutic efficiency.However,the existing materials often have the disadvantages of complicated and slow regeneration of blood vessels.Therefore,finding a scaffold with simple and convenient clinical availability is important.ECM plays an important regulatory role in cell proliferation,differentiation and gene expression,and has 3D structure of natural tissue.It has been widely used in tissue engineering.DAT,an extracellular matrix derived from adipose tissue,has low immunogenicity and can promote adipose tissue neovascularization and angiogenesis.The combined transplantation of DAT and ADSCs will provide new ideas for the repair and reconstruction of breast defects.Objective: To study ADSCs as seed cells,DAT as scaffold,and the DAT-ADSCs graft were co-cultured in vitro.Investigating the effects of DAT 3D scaffold on the biological characteristics of ADSCs,and the feasibility and advantages of DAT as the scaffold material;systematically compareing the effects of 2D environment and DAT on adipogenic differentiation of ADSCs,observing the outcomes of DAT-ADSCs grafts,and conducting initial exploration for the construction of engineering adipose tissue.Methods: Explore a simple and feasible DAT method and determine its biological shape and safety;To separate and culture ADSCs,phenotypic identification and adipogenic induction of P3 generation were performed to determine their adipogenic differentiation ability;To compare the biological characteristics of ADSCs cultured in 2D and DAT 3D culture,to determine the adhesion and proliferation rate of ADSCs under DAT culture,to explore the effect of DAT on adipogenic induction of ADSCs.Results: 1.The primary cultured ADSCs can reach 70%-80% fusion in 7-9 days.After subculture,the plastic adherent ADSCs show spindle-shaped and fibroblast-like cells.After 2 weeks,the cells gradually changed from shuttle to elliptical,and merged from smaller lipid droplets into larger ones.The lipid droplets in the cytoplasm were orange-red after oil red O staining.2.Per 400 ml liposuction can be prepared to approximately 15-30 ml of DAT,which is a milky white gel-like structure.DAPI fluorescence staining showed the cell-free components of the DAT remained.The histological staining results showed the DAT was mainly collagen.SEM showed DAT powder have a larger surface area and a higher porosity.3.The adhesion rate of ADSCs in the DAT was 86%.The proliferation rate was detected by MTT assay.The results showed that ADSCs could achieve good adhesion and no cytotoxicity in DAT scaffolds.The proliferation ability of ADSCs was not affected.ADSCs combined with DAT can induce ADSCs into mature adipocytes after 14 days in vitro.Oil red O staining shows DAT can induce ADSCs into matrue adipocytes.Conclusion: 1.ADSCs are widely sourced and easily isolated.ADSCs have the potential to differentiate into mature adipose tissue.2.The DAT is easy to obtain and it can be processed into various shapes.It removes the cellular components thoroughly,and can preserve relatively intact extracellular matrix components,and has a good 3d structure and biological properties.3.When ADSCs are co-cultured with DAT,it can achieve good adhesion and stable proliferation,and differentiate into mature adipose tissue under appropriate induction conditions.In addition,DAT can be used as an ideal scaffold material for adipose tissue engineering.